EP1558922B1 - Identifying therapeutic compounds based on their physical-chemical properties - Google Patents
Identifying therapeutic compounds based on their physical-chemical properties Download PDFInfo
- Publication number
- EP1558922B1 EP1558922B1 EP03781483A EP03781483A EP1558922B1 EP 1558922 B1 EP1558922 B1 EP 1558922B1 EP 03781483 A EP03781483 A EP 03781483A EP 03781483 A EP03781483 A EP 03781483A EP 1558922 B1 EP1558922 B1 EP 1558922B1
- Authority
- EP
- European Patent Office
- Prior art keywords
- potential
- physical
- parameter
- oxidation
- compounds
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 150000001875 compounds Chemical class 0.000 title claims description 101
- 239000000126 substance Substances 0.000 title claims description 60
- 230000001225 therapeutic effect Effects 0.000 title claims description 14
- 230000003647 oxidation Effects 0.000 claims description 74
- 238000007254 oxidation reaction Methods 0.000 claims description 74
- 238000000034 method Methods 0.000 claims description 64
- 230000009467 reduction Effects 0.000 claims description 51
- 238000003556 assay Methods 0.000 claims description 32
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 28
- 208000028867 ischemia Diseases 0.000 claims description 21
- 229910021607 Silver chloride Inorganic materials 0.000 claims description 20
- 229910052709 silver Inorganic materials 0.000 claims description 20
- 239000004332 silver Substances 0.000 claims description 20
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 claims description 20
- 208000035475 disorder Diseases 0.000 claims description 18
- 230000004770 neurodegeneration Effects 0.000 claims description 14
- 238000012360 testing method Methods 0.000 claims description 14
- 230000004071 biological effect Effects 0.000 claims description 13
- 230000004054 inflammatory process Effects 0.000 claims description 12
- 230000036542 oxidative stress Effects 0.000 claims description 12
- 206010021143 Hypoxia Diseases 0.000 claims description 11
- 206010061218 Inflammation Diseases 0.000 claims description 11
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 11
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 claims description 9
- 238000001075 voltammogram Methods 0.000 claims description 8
- VMWNQDUVQKEIOC-CYBMUJFWSA-N apomorphine Chemical group C([C@H]1N(C)CC2)C3=CC=C(O)C(O)=C3C3=C1C2=CC=C3 VMWNQDUVQKEIOC-CYBMUJFWSA-N 0.000 claims description 7
- 229930195712 glutamate Natural products 0.000 claims description 7
- PJANXHGTPQOBST-UHFFFAOYSA-N stilbene Chemical group C=1C=CC=CC=1C=CC1=CC=CC=C1 PJANXHGTPQOBST-UHFFFAOYSA-N 0.000 claims description 7
- 238000006467 substitution reaction Methods 0.000 claims description 7
- 208000024827 Alzheimer disease Diseases 0.000 claims description 6
- 230000015572 biosynthetic process Effects 0.000 claims description 6
- 230000030833 cell death Effects 0.000 claims description 6
- 230000000971 hippocampal effect Effects 0.000 claims description 6
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 6
- 238000000423 cell based assay Methods 0.000 claims description 5
- 125000004151 quinonyl group Chemical group 0.000 claims description 5
- 102000015689 E-Selectin Human genes 0.000 claims description 4
- 108010024212 E-Selectin Proteins 0.000 claims description 4
- 238000000159 protein binding assay Methods 0.000 claims description 4
- JADVWWSKYZXRGX-UHFFFAOYSA-M thioflavine T Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C1=[N+](C)C2=CC=C(C)C=C2S1 JADVWWSKYZXRGX-UHFFFAOYSA-M 0.000 claims description 4
- 206010002660 Anoxia Diseases 0.000 claims description 3
- 229940124598 therapeutic candidate Drugs 0.000 claims description 3
- 238000010822 cell death assay Methods 0.000 claims description 2
- 102100023471 E-selectin Human genes 0.000 claims 3
- 102000013455 Amyloid beta-Peptides Human genes 0.000 claims 1
- 108010090849 Amyloid beta-Peptides Proteins 0.000 claims 1
- 241000976983 Anoxia Species 0.000 claims 1
- 102000003800 Selectins Human genes 0.000 claims 1
- 108090000184 Selectins Proteins 0.000 claims 1
- 230000007953 anoxia Effects 0.000 claims 1
- VZWXIQHBIQLMPN-UHFFFAOYSA-N chromane Chemical group C1=CC=C2CCCOC2=C1 VZWXIQHBIQLMPN-UHFFFAOYSA-N 0.000 claims 1
- 125000004122 cyclic group Chemical group 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 46
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 42
- 239000000243 solution Substances 0.000 description 38
- 239000000203 mixture Substances 0.000 description 36
- 230000000694 effects Effects 0.000 description 24
- 210000002569 neuron Anatomy 0.000 description 24
- 238000002484 cyclic voltammetry Methods 0.000 description 19
- 239000012981 Hank's balanced salt solution Substances 0.000 description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 16
- 230000001537 neural effect Effects 0.000 description 14
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 12
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 12
- 238000005259 measurement Methods 0.000 description 11
- 238000006243 chemical reaction Methods 0.000 description 10
- 201000010099 disease Diseases 0.000 description 10
- 239000002609 medium Substances 0.000 description 10
- 239000000546 pharmaceutical excipient Substances 0.000 description 10
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 10
- 239000000725 suspension Substances 0.000 description 10
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 9
- 238000009472 formulation Methods 0.000 description 9
- 239000008103 glucose Substances 0.000 description 9
- 238000002751 oxidative stress assay Methods 0.000 description 9
- 238000004365 square wave voltammetry Methods 0.000 description 9
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 8
- 238000010521 absorption reaction Methods 0.000 description 8
- 150000002215 flavonoids Chemical class 0.000 description 8
- 238000002360 preparation method Methods 0.000 description 8
- 230000008569 process Effects 0.000 description 8
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 8
- 208000024891 symptom Diseases 0.000 description 8
- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 7
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 7
- 239000000872 buffer Substances 0.000 description 7
- 239000002552 dosage form Substances 0.000 description 7
- 239000001301 oxygen Substances 0.000 description 7
- 229910052760 oxygen Inorganic materials 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 239000011780 sodium chloride Substances 0.000 description 7
- 125000001424 substituent group Chemical group 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 208000006011 Stroke Diseases 0.000 description 6
- 239000004480 active ingredient Substances 0.000 description 6
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 6
- 210000004556 brain Anatomy 0.000 description 6
- 230000000875 corresponding effect Effects 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 229940049906 glutamate Drugs 0.000 description 6
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 6
- 208000015122 neurodegenerative disease Diseases 0.000 description 6
- 239000003642 reactive oxygen metabolite Substances 0.000 description 6
- 230000002829 reductive effect Effects 0.000 description 6
- 241000894007 species Species 0.000 description 6
- IPJDHSYCSQAODE-UHFFFAOYSA-N 5-chloromethylfluorescein diacetate Chemical compound O1C(=O)C2=CC(CCl)=CC=C2C21C1=CC=C(OC(C)=O)C=C1OC1=CC(OC(=O)C)=CC=C21 IPJDHSYCSQAODE-UHFFFAOYSA-N 0.000 description 5
- 238000005481 NMR spectroscopy Methods 0.000 description 5
- 230000000903 blocking effect Effects 0.000 description 5
- 230000001419 dependent effect Effects 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 210000002889 endothelial cell Anatomy 0.000 description 5
- 229930003935 flavonoid Natural products 0.000 description 5
- 235000017173 flavonoids Nutrition 0.000 description 5
- -1 flavonoids compounds Chemical class 0.000 description 5
- 231100000252 nontoxic Toxicity 0.000 description 5
- 230000003000 nontoxic effect Effects 0.000 description 5
- 230000001590 oxidative effect Effects 0.000 description 5
- 239000008194 pharmaceutical composition Substances 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- KDXKERNSBIXSRK-RXMQYKEDSA-N D-lysine Chemical compound NCCCC[C@@H](N)C(O)=O KDXKERNSBIXSRK-RXMQYKEDSA-N 0.000 description 4
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 4
- 229930182816 L-glutamine Natural products 0.000 description 4
- 229930182555 Penicillin Natural products 0.000 description 4
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 4
- REFJWTPEDVJJIY-UHFFFAOYSA-N Quercetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 REFJWTPEDVJJIY-UHFFFAOYSA-N 0.000 description 4
- 238000013459 approach Methods 0.000 description 4
- 229910052786 argon Inorganic materials 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 238000002848 electrochemical method Methods 0.000 description 4
- 230000001146 hypoxic effect Effects 0.000 description 4
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 description 4
- 229940049954 penicillin Drugs 0.000 description 4
- 229910052697 platinum Inorganic materials 0.000 description 4
- 238000010926 purge Methods 0.000 description 4
- 229960001285 quercetin Drugs 0.000 description 4
- 230000002468 redox effect Effects 0.000 description 4
- 230000027756 respiratory electron transport chain Effects 0.000 description 4
- 238000012552 review Methods 0.000 description 4
- 239000002002 slurry Substances 0.000 description 4
- 229960005322 streptomycin Drugs 0.000 description 4
- 238000005556 structure-activity relationship Methods 0.000 description 4
- 238000013268 sustained release Methods 0.000 description 4
- 208000011580 syndromic disease Diseases 0.000 description 4
- 239000003826 tablet Substances 0.000 description 4
- KBLZDCFTQSIIOH-UHFFFAOYSA-M tetrabutylazanium;perchlorate Chemical compound [O-]Cl(=O)(=O)=O.CCCC[N+](CCCC)(CCCC)CCCC KBLZDCFTQSIIOH-UHFFFAOYSA-M 0.000 description 4
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 3
- 239000012583 B-27 Supplement Substances 0.000 description 3
- UHDGCWIWMRVCDJ-CCXZUQQUSA-N Cytarabine Chemical compound O=C1N=C(N)C=CN1[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O1 UHDGCWIWMRVCDJ-CCXZUQQUSA-N 0.000 description 3
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 3
- ZVOLCUVKHLEPEV-UHFFFAOYSA-N Quercetagetin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=C(O)C(O)=C(O)C=C2O1 ZVOLCUVKHLEPEV-UHFFFAOYSA-N 0.000 description 3
- 241000700159 Rattus Species 0.000 description 3
- HWTZYBCRDDUBJY-UHFFFAOYSA-N Rhynchosin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=CC(O)=C(O)C=C2O1 HWTZYBCRDDUBJY-UHFFFAOYSA-N 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 229930006000 Sucrose Natural products 0.000 description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 229940114079 arachidonic acid Drugs 0.000 description 3
- 235000021342 arachidonic acid Nutrition 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 230000005779 cell damage Effects 0.000 description 3
- 230000003833 cell viability Effects 0.000 description 3
- 239000001913 cellulose Substances 0.000 description 3
- 229920002678 cellulose Polymers 0.000 description 3
- 235000010980 cellulose Nutrition 0.000 description 3
- 210000003169 central nervous system Anatomy 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000009792 diffusion process Methods 0.000 description 3
- 230000003291 dopaminomimetic effect Effects 0.000 description 3
- 239000002158 endotoxin Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 229960003180 glutathione Drugs 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000013537 high throughput screening Methods 0.000 description 3
- 210000001320 hippocampus Anatomy 0.000 description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 3
- 230000001965 increasing effect Effects 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000008101 lactose Substances 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- 229920006008 lipopolysaccharide Polymers 0.000 description 3
- 235000019359 magnesium stearate Nutrition 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 239000006187 pill Substances 0.000 description 3
- 239000013641 positive control Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 235000005875 quercetin Nutrition 0.000 description 3
- 238000010405 reoxidation reaction Methods 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 230000035882 stress Effects 0.000 description 3
- 239000005720 sucrose Substances 0.000 description 3
- 239000012730 sustained-release form Substances 0.000 description 3
- 239000000454 talc Substances 0.000 description 3
- 235000012222 talc Nutrition 0.000 description 3
- 229910052623 talc Inorganic materials 0.000 description 3
- 239000012085 test solution Substances 0.000 description 3
- DZUOQMBJJSBONO-CQSZACIVSA-N (6ar)-10-methoxy-6-methyl-5,6,6a,7-tetrahydro-4h-dibenzo[de,g]quinoline-11-ol Chemical compound CN1CCC2=CC=CC3=C2[C@H]1CC1=CC=C(OC)C(O)=C13 DZUOQMBJJSBONO-CQSZACIVSA-N 0.000 description 2
- ICLYJLBTOGPLMC-KVVVOXFISA-N (z)-octadec-9-enoate;tris(2-hydroxyethyl)azanium Chemical compound OCCN(CCO)CCO.CCCCCCCC\C=C/CCCCCCCC(O)=O ICLYJLBTOGPLMC-KVVVOXFISA-N 0.000 description 2
- WOAHJDHKFWSLKE-UHFFFAOYSA-N 1,2-benzoquinone Chemical group O=C1C=CC=CC1=O WOAHJDHKFWSLKE-UHFFFAOYSA-N 0.000 description 2
- KBPLFHHGFOOTCA-UHFFFAOYSA-N 1-Octanol Chemical compound CCCCCCCCO KBPLFHHGFOOTCA-UHFFFAOYSA-N 0.000 description 2
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 2
- XZIIFPSPUDAGJM-UHFFFAOYSA-N 6-chloro-2-n,2-n-diethylpyrimidine-2,4-diamine Chemical compound CCN(CC)C1=NC(N)=CC(Cl)=N1 XZIIFPSPUDAGJM-UHFFFAOYSA-N 0.000 description 2
- 208000002874 Acne Vulgaris Diseases 0.000 description 2
- 206010001052 Acute respiratory distress syndrome Diseases 0.000 description 2
- 208000023275 Autoimmune disease Diseases 0.000 description 2
- 201000006474 Brain Ischemia Diseases 0.000 description 2
- 108010074051 C-Reactive Protein Proteins 0.000 description 2
- 102100032752 C-reactive protein Human genes 0.000 description 2
- 206010008089 Cerebral artery occlusion Diseases 0.000 description 2
- 206010008120 Cerebral ischaemia Diseases 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 201000004624 Dermatitis Diseases 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- 102000018899 Glutamate Receptors Human genes 0.000 description 2
- 108010027915 Glutamate Receptors Proteins 0.000 description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- 108010024636 Glutathione Proteins 0.000 description 2
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 2
- 208000010496 Heart Arrest Diseases 0.000 description 2
- 208000032087 Hereditary Leber Optic Atrophy Diseases 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 201000000639 Leber hereditary optic neuropathy Diseases 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- HOKKHZGPKSLGJE-GSVOUGTGSA-N N-Methyl-D-aspartic acid Chemical compound CN[C@@H](C(O)=O)CC(O)=O HOKKHZGPKSLGJE-GSVOUGTGSA-N 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- 208000024777 Prion disease Diseases 0.000 description 2
- 208000013616 Respiratory Distress Syndrome Diseases 0.000 description 2
- 206010039792 Seborrhoea Diseases 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- PJANXHGTPQOBST-VAWYXSNFSA-N Stilbene Natural products C=1C=CC=CC=1/C=C/C1=CC=CC=C1 PJANXHGTPQOBST-VAWYXSNFSA-N 0.000 description 2
- 239000004809 Teflon Substances 0.000 description 2
- 229920006362 Teflon® Polymers 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 206010000496 acne Diseases 0.000 description 2
- 208000011341 adult acute respiratory distress syndrome Diseases 0.000 description 2
- 201000000028 adult respiratory distress syndrome Diseases 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 206010064930 age-related macular degeneration Diseases 0.000 description 2
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 2
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 description 2
- 239000012491 analyte Substances 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 229960004046 apomorphine Drugs 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 239000012298 atmosphere Substances 0.000 description 2
- 201000008937 atopic dermatitis Diseases 0.000 description 2
- 208000010668 atopic eczema Diseases 0.000 description 2
- 230000006931 brain damage Effects 0.000 description 2
- 231100000874 brain damage Toxicity 0.000 description 2
- 208000029028 brain injury Diseases 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 208000037887 cell injury Diseases 0.000 description 2
- 206010008118 cerebral infarction Diseases 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 231100000433 cytotoxic Toxicity 0.000 description 2
- 230000001472 cytotoxic effect Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 239000008121 dextrose Substances 0.000 description 2
- 229910003460 diamond Inorganic materials 0.000 description 2
- 239000010432 diamond Substances 0.000 description 2
- VAGAHQGPIKRLLK-UHFFFAOYSA-N diplamine Chemical compound C1=NC(C(C(SC)=C2CCNC(C)=O)=O)=C3C2=NC2=CC=CC=C2C3=C1 VAGAHQGPIKRLLK-UHFFFAOYSA-N 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000003411 electrode reaction Methods 0.000 description 2
- 239000003995 emulsifying agent Substances 0.000 description 2
- 230000001815 facial effect Effects 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 238000005187 foaming Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 239000007903 gelatin capsule Substances 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 229910021397 glassy carbon Inorganic materials 0.000 description 2
- 150000002334 glycols Chemical class 0.000 description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- 229910052737 gold Inorganic materials 0.000 description 2
- 239000010931 gold Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 238000004770 highest occupied molecular orbital Methods 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 230000007954 hypoxia Effects 0.000 description 2
- 238000010874 in vitro model Methods 0.000 description 2
- 238000010348 incorporation Methods 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 239000000543 intermediate Substances 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 238000001990 intravenous administration Methods 0.000 description 2
- 230000000302 ischemic effect Effects 0.000 description 2
- 238000004768 lowest unoccupied molecular orbital Methods 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 208000002780 macular degeneration Diseases 0.000 description 2
- ZLNQQNXFFQJAID-UHFFFAOYSA-L magnesium carbonate Chemical compound [Mg+2].[O-]C([O-])=O ZLNQQNXFFQJAID-UHFFFAOYSA-L 0.000 description 2
- 239000001095 magnesium carbonate Substances 0.000 description 2
- 229910000021 magnesium carbonate Inorganic materials 0.000 description 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 201000007309 middle cerebral artery infarction Diseases 0.000 description 2
- 239000003068 molecular probe Substances 0.000 description 2
- 208000031225 myocardial ischemia Diseases 0.000 description 2
- 229920001778 nylon Polymers 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 235000019198 oils Nutrition 0.000 description 2
- 239000006179 pH buffering agent Substances 0.000 description 2
- 238000007911 parenteral administration Methods 0.000 description 2
- 238000005192 partition Methods 0.000 description 2
- 230000035699 permeability Effects 0.000 description 2
- 238000000053 physical method Methods 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 238000005498 polishing Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- RUOJZAUFBMNUDX-UHFFFAOYSA-N propylene carbonate Chemical compound CC1COC(=O)O1 RUOJZAUFBMNUDX-UHFFFAOYSA-N 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 2
- 239000012266 salt solution Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 229940035044 sorbitan monolaurate Drugs 0.000 description 2
- 235000021286 stilbenes Nutrition 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000003950 stripping voltammetry Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000000699 topical effect Effects 0.000 description 2
- 150000003626 triacylglycerols Chemical class 0.000 description 2
- 229940117013 triethanolamine oleate Drugs 0.000 description 2
- 235000015112 vegetable and seed oil Nutrition 0.000 description 2
- 239000008158 vegetable oil Substances 0.000 description 2
- 238000004832 voltammetry Methods 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- 239000001100 (2S)-5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl)chroman-4-one Substances 0.000 description 1
- MHPQCGZBAVXCGA-GFCCVEGCSA-N (6ar)-5,6,6a,7-tetrahydro-4h-dibenzo[de,g]quinoline-10,11-diol Chemical compound N1CCC2=CC=CC3=C2[C@H]1CC1=CC=C(O)C(O)=C13 MHPQCGZBAVXCGA-GFCCVEGCSA-N 0.000 description 1
- BTGAJCKRXPNBFI-OAHLLOKOSA-N (8R)-7-propyl-5,6,6a,7-tetrahydro-4H-dibenzo[de,g]quinoline-13,14-diol Chemical compound C([C@H]1N(CCC)CC2)C3=CC=C(O)C(O)=C3C3=C1C2=CC=C3 BTGAJCKRXPNBFI-OAHLLOKOSA-N 0.000 description 1
- 150000000191 1,4-naphthoquinones Chemical class 0.000 description 1
- BTLXPCBPYBNQNR-UHFFFAOYSA-N 1-hydroxyanthraquinone Chemical class O=C1C2=CC=CC=C2C(=O)C2=C1C=CC=C2O BTLXPCBPYBNQNR-UHFFFAOYSA-N 0.000 description 1
- RZVHIXYEVGDQDX-UHFFFAOYSA-N 9,10-anthraquinone Chemical group C1=CC=C2C(=O)C3=CC=CC=C3C(=O)C2=C1 RZVHIXYEVGDQDX-UHFFFAOYSA-N 0.000 description 1
- 206010065040 AIDS dementia complex Diseases 0.000 description 1
- 244000215068 Acacia senegal Species 0.000 description 1
- 235000006491 Acacia senegal Nutrition 0.000 description 1
- 206010049001 Acute endocarditis Diseases 0.000 description 1
- 201000004384 Alopecia Diseases 0.000 description 1
- 206010002023 Amyloidoses Diseases 0.000 description 1
- 102000011730 Arachidonate 12-Lipoxygenase Human genes 0.000 description 1
- 108010076676 Arachidonate 12-lipoxygenase Proteins 0.000 description 1
- 206010003591 Ataxia Diseases 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 206010004446 Benign prostatic hyperplasia Diseases 0.000 description 1
- 208000002381 Brain Hypoxia Diseases 0.000 description 1
- 206010066091 Bronchial Hyperreactivity Diseases 0.000 description 1
- LODGIKWNLDQZBM-UHFFFAOYSA-N Bulbocapnine Natural products C12=C3C4=C(O)C(OC)=CC=C4CC2N(C)CCC1=CC1=C3OCO1 LODGIKWNLDQZBM-UHFFFAOYSA-N 0.000 description 1
- 206010007558 Cardiac failure chronic Diseases 0.000 description 1
- 206010007559 Cardiac failure congestive Diseases 0.000 description 1
- 208000031229 Cardiomyopathies Diseases 0.000 description 1
- 102000016289 Cell Adhesion Molecules Human genes 0.000 description 1
- 108010067225 Cell Adhesion Molecules Proteins 0.000 description 1
- 208000035484 Cellulite Diseases 0.000 description 1
- 208000005145 Cerebral amyloid angiopathy Diseases 0.000 description 1
- 201000006082 Chickenpox Diseases 0.000 description 1
- 208000006545 Chronic Obstructive Pulmonary Disease Diseases 0.000 description 1
- 206010009900 Colitis ulcerative Diseases 0.000 description 1
- 208000020406 Creutzfeldt Jacob disease Diseases 0.000 description 1
- 208000003407 Creutzfeldt-Jakob Syndrome Diseases 0.000 description 1
- 208000010859 Creutzfeldt-Jakob disease Diseases 0.000 description 1
- 208000011231 Crohn disease Diseases 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- 206010012434 Dermatitis allergic Diseases 0.000 description 1
- 206010012438 Dermatitis atopic Diseases 0.000 description 1
- 206010012442 Dermatitis contact Diseases 0.000 description 1
- 208000002249 Diabetes Complications Diseases 0.000 description 1
- 208000032131 Diabetic Neuropathies Diseases 0.000 description 1
- 206010056340 Diabetic ulcer Diseases 0.000 description 1
- 235000019739 Dicalciumphosphate Nutrition 0.000 description 1
- 201000010374 Down Syndrome Diseases 0.000 description 1
- 206010013801 Duchenne Muscular Dystrophy Diseases 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 208000024412 Friedreich ataxia Diseases 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- 206010019196 Head injury Diseases 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 206010056328 Hepatic ischaemia Diseases 0.000 description 1
- 208000023105 Huntington disease Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010020880 Hypertrophy Diseases 0.000 description 1
- 208000001953 Hypotension Diseases 0.000 description 1
- 208000022559 Inflammatory bowel disease Diseases 0.000 description 1
- 206010022680 Intestinal ischaemia Diseases 0.000 description 1
- QELDJEKNFOQJOY-ZDUSSCGKSA-N Isocorydine Chemical compound CN1CCC2=CC(OC)=C(OC)C3=C2[C@@H]1CC1=CC=C(OC)C(O)=C13 QELDJEKNFOQJOY-ZDUSSCGKSA-N 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- LEVWYRKDKASIDU-IMJSIDKUSA-N L-cystine Chemical compound [O-]C(=O)[C@@H]([NH3+])CSSC[C@H]([NH3+])C([O-])=O LEVWYRKDKASIDU-IMJSIDKUSA-N 0.000 description 1
- 208000019693 Lung disease Diseases 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 201000009906 Meningitis Diseases 0.000 description 1
- 229930192392 Mitomycin Natural products 0.000 description 1
- 239000004909 Moisturizer Substances 0.000 description 1
- 206010049565 Muscle fatigue Diseases 0.000 description 1
- 208000009525 Myocarditis Diseases 0.000 description 1
- 240000001307 Myosotis scorpioides Species 0.000 description 1
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000028389 Nerve injury Diseases 0.000 description 1
- 206010029333 Neurosis Diseases 0.000 description 1
- GZCJJOLJSBCUNR-UHFFFAOYSA-N Oc(cc1)cc2c1OCCC2 Chemical compound Oc(cc1)cc2c1OCCC2 GZCJJOLJSBCUNR-UHFFFAOYSA-N 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 208000005141 Otitis Diseases 0.000 description 1
- 206010033892 Paraplegia Diseases 0.000 description 1
- 208000018737 Parkinson disease Diseases 0.000 description 1
- 206010049752 Peau d'orange Diseases 0.000 description 1
- 206010051246 Photodermatosis Diseases 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 241000241413 Propolis Species 0.000 description 1
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical class CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000004403 Prostatic Hyperplasia Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 208000003251 Pruritus Diseases 0.000 description 1
- 201000004681 Psoriasis Diseases 0.000 description 1
- 206010063562 Radiation skin injury Diseases 0.000 description 1
- 206010063897 Renal ischaemia Diseases 0.000 description 1
- 206010063837 Reperfusion injury Diseases 0.000 description 1
- 208000017442 Retinal disease Diseases 0.000 description 1
- 206010038923 Retinopathy Diseases 0.000 description 1
- 241001303601 Rosacea Species 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- 206010039966 Senile dementia Diseases 0.000 description 1
- 208000021386 Sjogren Syndrome Diseases 0.000 description 1
- 206010040799 Skin atrophy Diseases 0.000 description 1
- 206010040925 Skin striae Diseases 0.000 description 1
- 206010040954 Skin wrinkling Diseases 0.000 description 1
- 208000036982 Spinal cord ischaemia Diseases 0.000 description 1
- 208000031439 Striae Distensae Diseases 0.000 description 1
- 208000022292 Tay-Sachs disease Diseases 0.000 description 1
- 206010053615 Thermal burn Diseases 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 241000159243 Toxicodendron radicans Species 0.000 description 1
- 208000032109 Transient ischaemic attack Diseases 0.000 description 1
- 206010044688 Trisomy 21 Diseases 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 206010067584 Type 1 diabetes mellitus Diseases 0.000 description 1
- 201000006704 Ulcerative Colitis Diseases 0.000 description 1
- 206010046980 Varicella Diseases 0.000 description 1
- 206010052428 Wound Diseases 0.000 description 1
- 206010048222 Xerosis Diseases 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000000370 acceptor Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 150000001447 alkali salts Chemical class 0.000 description 1
- 231100000360 alopecia Toxicity 0.000 description 1
- VREFGVBLTWBCJP-UHFFFAOYSA-N alprazolam Chemical compound C12=CC(Cl)=CC=C2N2C(C)=NN=C2CN=C1C1=CC=CC=C1 VREFGVBLTWBCJP-UHFFFAOYSA-N 0.000 description 1
- 206010002022 amyloidosis Diseases 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 229930014669 anthocyanidin Natural products 0.000 description 1
- 150000001452 anthocyanidin derivatives Chemical class 0.000 description 1
- 235000008758 anthocyanidins Nutrition 0.000 description 1
- 229940045799 anthracyclines and related substance Drugs 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 238000003782 apoptosis assay Methods 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 230000003190 augmentative effect Effects 0.000 description 1
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- XXJFCCXVBKGMRM-UHFFFAOYSA-N benzo[b][4,7]phenanthroline Chemical class C1=CC=C2C3=CC4=CC=CC=C4N=C3C=CC2=N1 XXJFCCXVBKGMRM-UHFFFAOYSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000036427 bronchial hyperreactivity Effects 0.000 description 1
- LODGIKWNLDQZBM-LBPRGKRZSA-N bulbocapnine Chemical compound CN([C@H]1CC2=CC=C(C(=C2C2=C11)O)OC)CCC1=CC1=C2OCO1 LODGIKWNLDQZBM-LBPRGKRZSA-N 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- LLSDKQJKOVVTOJ-UHFFFAOYSA-L calcium chloride dihydrate Chemical compound O.O.[Cl-].[Cl-].[Ca+2] LLSDKQJKOVVTOJ-UHFFFAOYSA-L 0.000 description 1
- 230000009460 calcium influx Effects 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000017455 cell-cell adhesion Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000036232 cellulite Effects 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 238000001567 chrono-coulometry Methods 0.000 description 1
- 238000004769 chrono-potentiometry Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000004590 computer program Methods 0.000 description 1
- 239000004020 conductor Substances 0.000 description 1
- 208000010247 contact dermatitis Diseases 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 239000012059 conventional drug carrier Substances 0.000 description 1
- 210000004351 coronary vessel Anatomy 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 229940097362 cyclodextrins Drugs 0.000 description 1
- 229960003067 cystine Drugs 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000003412 degenerative effect Effects 0.000 description 1
- 230000036576 dermal application Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- NEFBYIFKOOEVPA-UHFFFAOYSA-K dicalcium phosphate Chemical compound [Ca+2].[Ca+2].[O-]P([O-])([O-])=O NEFBYIFKOOEVPA-UHFFFAOYSA-K 0.000 description 1
- 229940038472 dicalcium phosphate Drugs 0.000 description 1
- 229910000390 dicalcium phosphate Inorganic materials 0.000 description 1
- 150000005690 diesters Chemical class 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 229930190788 diplamine Natural products 0.000 description 1
- 239000007919 dispersible tablet Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 210000004002 dopaminergic cell Anatomy 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 239000000890 drug combination Substances 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 238000003487 electrochemical reaction Methods 0.000 description 1
- 230000005672 electromagnetic field Effects 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 206010014599 encephalitis Diseases 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 206010015037 epilepsy Diseases 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000003492 excitotoxic effect Effects 0.000 description 1
- 231100000063 excitotoxicity Toxicity 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000002795 fluorescence method Methods 0.000 description 1
- IAODRFIZLKITMK-UHFFFAOYSA-N furan-2,3-dione Chemical class O=C1OC=CC1=O IAODRFIZLKITMK-UHFFFAOYSA-N 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 208000007565 gingivitis Diseases 0.000 description 1
- 230000007946 glucose deprivation Effects 0.000 description 1
- 230000004190 glucose uptake Effects 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 230000009931 harmful effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000010224 hepatic metabolism Effects 0.000 description 1
- AIONOLUJZLIMTK-UHFFFAOYSA-N hesperetin Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(O)=CC(O)=C2C(=O)C1 AIONOLUJZLIMTK-UHFFFAOYSA-N 0.000 description 1
- AIONOLUJZLIMTK-AWEZNQCLSA-N hesperetin Chemical compound C1=C(O)C(OC)=CC=C1[C@H]1OC2=CC(O)=CC(O)=C2C(=O)C1 AIONOLUJZLIMTK-AWEZNQCLSA-N 0.000 description 1
- 229960001587 hesperetin Drugs 0.000 description 1
- 235000010209 hesperetin Nutrition 0.000 description 1
- 210000004295 hippocampal neuron Anatomy 0.000 description 1
- FTODBIPDTXRIGS-UHFFFAOYSA-N homoeriodictyol Natural products C1=C(O)C(OC)=CC(C2OC3=CC(O)=CC(O)=C3C(=O)C2)=C1 FTODBIPDTXRIGS-UHFFFAOYSA-N 0.000 description 1
- 150000004420 hydroxy-1,4-naphthoquinones Chemical class 0.000 description 1
- 230000036543 hypotension Effects 0.000 description 1
- 208000018875 hypoxemia Diseases 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000005462 in vivo assay Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000004968 inflammatory condition Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000297 inotrophic effect Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000007913 intrathecal administration Methods 0.000 description 1
- 238000007914 intraventricular administration Methods 0.000 description 1
- DRVVGOFZCWCCSA-UHFFFAOYSA-N isocorydine Natural products COc1cc2CCN(C)C3Cc4cccc(C)c4c(c1O)c23 DRVVGOFZCWCCSA-UHFFFAOYSA-N 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000003780 keratinization Effects 0.000 description 1
- 201000010901 lateral sclerosis Diseases 0.000 description 1
- 230000023404 leukocyte cell-cell adhesion Effects 0.000 description 1
- 239000008297 liquid dosage form Substances 0.000 description 1
- 239000006193 liquid solution Substances 0.000 description 1
- 239000006194 liquid suspension Substances 0.000 description 1
- 230000027928 long-term synaptic potentiation Effects 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- LRDGATPGVJTWLJ-UHFFFAOYSA-N luteolin Natural products OC1=CC(O)=CC(C=2OC3=CC(O)=CC(O)=C3C(=O)C=2)=C1 LRDGATPGVJTWLJ-UHFFFAOYSA-N 0.000 description 1
- IQPNAANSBPBGFQ-UHFFFAOYSA-N luteolin Chemical compound C=1C(O)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(O)C(O)=C1 IQPNAANSBPBGFQ-UHFFFAOYSA-N 0.000 description 1
- 235000009498 luteolin Nutrition 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 210000002418 meninge Anatomy 0.000 description 1
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 1
- 210000004925 microvascular endothelial cell Anatomy 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 230000005787 mitochondrial ATP synthesis coupled electron transport Effects 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000001333 moisturizer Effects 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- LPUQAYUQRXPFSQ-DFWYDOINSA-M monosodium L-glutamate Chemical compound [Na+].[O-]C(=O)[C@@H](N)CCC(O)=O LPUQAYUQRXPFSQ-DFWYDOINSA-M 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 208000005264 motor neuron disease Diseases 0.000 description 1
- 206010028417 myasthenia gravis Diseases 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 210000000107 myocyte Anatomy 0.000 description 1
- SYSQUGFVNFXIIT-UHFFFAOYSA-N n-[4-(1,3-benzoxazol-2-yl)phenyl]-4-nitrobenzenesulfonamide Chemical class C1=CC([N+](=O)[O-])=CC=C1S(=O)(=O)NC1=CC=C(C=2OC3=CC=CC=C3N=2)C=C1 SYSQUGFVNFXIIT-UHFFFAOYSA-N 0.000 description 1
- FRASJONUBLZVQX-UHFFFAOYSA-N naphthoquinone group Chemical group C1(C=CC(C2=CC=CC=C12)=O)=O FRASJONUBLZVQX-UHFFFAOYSA-N 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000008764 nerve damage Effects 0.000 description 1
- 208000004296 neuralgia Diseases 0.000 description 1
- 230000003955 neuronal function Effects 0.000 description 1
- 230000003961 neuronal insult Effects 0.000 description 1
- 230000007512 neuronal protection Effects 0.000 description 1
- 208000021722 neuropathic pain Diseases 0.000 description 1
- 201000001119 neuropathy Diseases 0.000 description 1
- 230000007823 neuropathy Effects 0.000 description 1
- 208000015238 neurotic disease Diseases 0.000 description 1
- 230000007959 normoxia Effects 0.000 description 1
- 230000037312 oily skin Effects 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 201000008482 osteoarthritis Diseases 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 208000008494 pericarditis Diseases 0.000 description 1
- 210000000578 peripheral nerve Anatomy 0.000 description 1
- 239000008177 pharmaceutical agent Substances 0.000 description 1
- 229940124531 pharmaceutical excipient Drugs 0.000 description 1
- 230000008845 photoaging Effects 0.000 description 1
- 230000008832 photodamage Effects 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 210000003281 pleural cavity Anatomy 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920006316 polyvinylpyrrolidine Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 230000003334 potential effect Effects 0.000 description 1
- 238000011533 pre-incubation Methods 0.000 description 1
- 210000005238 principal cell Anatomy 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000005522 programmed cell death Effects 0.000 description 1
- 210000001176 projection neuron Anatomy 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 229940069949 propolis Drugs 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 238000013139 quantization Methods 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 150000004059 quinone derivatives Chemical class 0.000 description 1
- 150000004053 quinones Chemical class 0.000 description 1
- 150000005838 radical anions Chemical class 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 238000006479 redox reaction Methods 0.000 description 1
- 230000006697 redox regulation Effects 0.000 description 1
- 230000010282 redox signaling Effects 0.000 description 1
- 238000011946 reduction process Methods 0.000 description 1
- 230000002207 retinal effect Effects 0.000 description 1
- 208000032253 retinal ischemia Diseases 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 201000004700 rosacea Diseases 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 230000037390 scarring Effects 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical class O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 208000008742 seborrheic dermatitis Diseases 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 230000009759 skin aging Effects 0.000 description 1
- 229960004249 sodium acetate Drugs 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 229940054269 sodium pyruvate Drugs 0.000 description 1
- 239000007909 solid dosage form Substances 0.000 description 1
- 239000008137 solubility enhancer Substances 0.000 description 1
- 208000020431 spinal cord injury Diseases 0.000 description 1
- 230000003393 splenic effect Effects 0.000 description 1
- 238000000141 square-wave voltammogram Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 230000003977 synaptic function Effects 0.000 description 1
- 230000003956 synaptic plasticity Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000007910 systemic administration Methods 0.000 description 1
- 201000000596 systemic lupus erythematosus Diseases 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000036962 time dependent Effects 0.000 description 1
- 238000011200 topical administration Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- QAIPRVGONGVQAS-DUXPYHPUSA-N trans-caffeic acid Chemical class OC(=O)\C=C\C1=CC=C(O)C(O)=C1 QAIPRVGONGVQAS-DUXPYHPUSA-N 0.000 description 1
- 238000006276 transfer reaction Methods 0.000 description 1
- 201000010875 transient cerebral ischemia Diseases 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- UPCXAARSWVHVLY-UHFFFAOYSA-N tris(2-hydroxyethyl)azanium;acetate Chemical compound CC(O)=O.OCCN(CCO)CCO UPCXAARSWVHVLY-UHFFFAOYSA-N 0.000 description 1
- 238000001665 trituration Methods 0.000 description 1
- 210000004291 uterus Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
- G01N33/6896—Neurological disorders, e.g. Alzheimer's disease
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/08—Bronchodilators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/08—Drugs for disorders of the urinary system of the prostate
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/08—Drugs for genital or sexual disorders; Contraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/08—Antiseborrheics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/12—Keratolytics, e.g. wart or anti-corn preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/14—Drugs for dermatological disorders for baldness or alopecia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/16—Emollients or protectives, e.g. against radiation
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
- A61P21/04—Drugs for disorders of the muscular or neuromuscular system for myasthenia gravis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/08—Antiepileptics; Anticonvulsants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
- A61P25/16—Anti-Parkinson drugs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2500/00—Screening for compounds of potential therapeutic value
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
Definitions
- the present invention is directed to novel methods for Identifying therapeutic compounds based on their structural and physlcal-chemical properties.
- novel drugs are Identified by screening compound libraries and determining which compounds have the highest therapeutic properties, and optimizing such properties by synthesizing structurally related analogs.
- the limitation of such an approach is that it is possible to synthesize and test only a very small subset of all possible molecules thereby resulting in a high probability that the most efficacious molecules will be missed.
- SAR activity by NMR as described In US 5,698,401 relates to a process for Identifying compounds which bind to a specific target molecule.
- the physical structure of a target protein is determined by nuclear magnetic resonance analysis (NMR) and the small molecule building blocks are Identified that bind to the protein at nearby points on the protein surface. Adjacently binding small molecules are then coupled together with a linker in order to obtain compounds that bind to the target protein with higher affinity that the unlinked compounds alone.
- NMR nuclear magnetic resonance analysis
- the "rule of 5" states that poor absorption or permeation are more likely when there are more than 5-H bond donors (expressed as the sum of OHs and NHs); more than 10 H-bond acceptors (expressed as the sum of Ns and Os); the molecular weight is over 500; and the log P is greater than 5.
- Cyclic voltammetry has been used for the detection of compounds In different solutions, see for example, Kilmartin, P. Antioxidants and Redox Signaling, 3, (6), 941-955, 2001 , and in a redox control and monitoring high throughput screening platform used In conjunction with another detector, (see US Application 2002/0123069 ), but heretofore cyclic voltammetry has not been used for the a priori identification of possible therapeutic candidates.
- the interaction of a molecule with a biological target may not be limited to structural recognition alone. Other types of interaction may occur, and some of them may involve the exchange of one or more charges. Because of their distinct electronic distribution, molecules exhibit different physical-chemical profiles, particularly redox profiles, i.e. the measure of an ability to give or accept charges, the number of charges exchanged, the kinetics of the process, and the subsequent molecular mechanisms that follow the addition or subtraction of charges. Various electroanalytical techniques can be employed to determine the physical-chemical profile. Cyclic voltammetry is a technique that yields quantitative information on the above parameters. The redox profile of a molecule is well characterized by its cyclic voltammogram.
- electroanalytical techniques may also be used to measure redox profile, such as for example single cyclic voltammetry, continuous cyclic voltammetry (with and without the integration of current), square wave voltammetry, square wave stripping voltammetry, AC voltammetry, choromoamperometry, chronocoulometry, chronopotentiometry, as well as various potentiostatic and galvanostatic techniques can be employed.
- a description of various electroanalytical techniques can be found in numerous textbooks, such as. Monk S, Fundamentals of Electroanalytical Chemistry, Wiley & Sons, New York,. 156-175, 2001 or Bard A.J. et al, Electrochemical Methods, Wiley & Sons, New York, Ch. 6, , 1980 .
- the above stated electroanalytical techniques should not be considered as limiting the scope of the invention, but merely as being illustrative and representative thereof.
- Cyclic voltammetry is a simple and direct method for measuring the formal potential of a half reaction when both oxidized and reduced forms are stable during the time required to obtain the current-potential curve.
- the resulting plot of current versus potential is termed a cyclic voltammogram.
- a cyclic voltammogram is a complicated, time-dependent function of a large number of physical and chemical parameters. It will give information on the anodic and cathodic peak potentials, the half-peak potential of the oxidation wave, the peak separation, the potential midway between the anodic and the cathodic peak. Only in the case where a redox system remains in equilibrium throughout the potential scan is the voltammogram reversible.
- the shape of a voltammogram can be significantly altered if there are coupled chemical reactions either before or after the electrochemical process.
- the mechanism, the rate, and the equilibrium constants of the process can all play a part in the final shape of the voltammogram that characterizes a certain molecule.
- Cyclic voltammetry is a powerful tool for the determination of formal redox potentials, detection of chemical reactions that precede or follow the electrochemical reaction, and evaluation of electron transfer kinetics.
- a cyclic voltammogram yields data on the ability of the molecule to shed one or more electrons measured by the oxidation potential, the relative number of electrons exchanged measured by the oxidation current, the reversibility of the oxidation process measured by the ratio of the oxidation current to the corresponding reduction current, and the kinetics of the oxidized species measured through variation of the scan rate; as well as the ability of the molecule to accept one or more electrons measured by the reduction potential, the number of electrons exchanged measured by the reduction current, the reversibility of the reduction process measured by the ratio of the reduction current to the corresponding oxidation current; and the stability of the reduced species measured through variation of the scan rate.
- Electrodes at which the competing redox reactions such as, for example, the hydrogen evolution reaction or oxygen evolution reaction, do not interfere.
- This can be achieved by employing electrodes characterized by sufficiently high overpotential towards the competing reactions, such as but not limited to platinum, gold and glassy carbon.
- the working electrode may be stationary or rotating, of any geometry, and can Include convective mixing.
- the measurements can be done in a variety of systems including but not limited to oxygenated or non-oxygenated environments, and protonated and non-protonated environments.
- the electro-chemical measurement effected herein is typically conducted in a three-electrode system (working electrode/counter electrode/reference electrode) or, possibly a two electrode system (working electrode/counter electrode).
- the area of the counter electrode may typically be larger than that of the measuring electrode.
- the energy profile of a molecule can be acquired from its optical values such as linear emission, linear absorption and non-linear absorption.
- Linear emission is measured by the wavelength of maximum fluorescence ⁇ em , i.e . the radiating energy released by the stabilization of an electron from a high energy level (LUMO) to a lower energy level (HOMO); and/or by quantum yield of fluorescence ⁇ , i.e . the probability of the molecule to stabilize an electron from its high energy level (LUMO) to a lower energy level (HOMO).
- Linear absorption is measured by the wavelength of maximum absorption ⁇ max, i.e .
- Polarizability ⁇ pol is measured by the ability of the electron cloud of the molecule to be shaped by an external electromagnetic field. Polarizability measurements are conducted by solvatochromism, measuring the shift in wavelength of maximum absorption as a function of solvent polarity (e.g. between octanol and DMSO).
- the transport profile of a molecule can be acquired from its partition coefficient, and/or diffusion constant, and/or molecular weight, and/or melting point.
- the biological activity of the small subset of compounds can be assessed by any assay deemed relevant to the target of interest.
- the biological activity of some of these compounds was determined using an in vitro model well recognized in the art for determining the degree of dysfunction of the cells when exposed to stress.
- In vitro models of ischemia approximate oxygen and glucose deprivation that mimic in vivo conditions, for example, by placing neuronal cultures into large anaerobic of hypoxic chambers and exchanging culture medium with de-oxygenated and defined ionic composition media.
- NMDA N-methyl-D-aspartate
- Oxidative stress has emerged as one of the major factors in the neurodegenerative disease and may contribute to neuronal damage from ischemia (see e.g., Coyle J. T. et al., Science 262, 689 -695, 1993 ; or Beal M. F, Curr. Opin. Neurobiol.; 6, 661-666, 1996 ).
- Reactive oxygen species ROS
- ROS reactive oxygen species
- HGOS glutarnate-Induced oxidative stress
- the cytotoxic effect of glutamate is not due to excitotoxicity, as this cell line is devoid of inotropic glutamate receptors. Rather, the glutamate-induced toxicity of dopaminergic cells is associated with an inhibition of cystine transport ( Murphy T. H., et al., Neuron 2,1547 -1558, 1989 ), which subsequently leads to depletion of intracellular glutathione (GSH) levels ( Murphy T. H., et al. Neuron 2,1547 -1558, 1989 ), activation of neuronal 12-lipoxygenase (U, Y.
- inflammation assays for example e-selectin (also named Endothelial Leukocyte Adhesion Molecule or ELAM) or C-reactive protein (CRP), or myocyte calcium-contractility assay or in vivo assays such as the rat middle cerebral artery occlusion (MCAO) model of cerebral ischemia assay, the rat paw edema assay or the mouse ear inflammation response to topical arachidonic acid assay or a skin protection assay may also be used.
- MCAO middle cerebral artery occlusion
- MCAO rat middle cerebral artery occlusion
- MCAO rat paw edema assay
- mouse ear inflammation response to topical arachidonic acid assay or a skin protection assay may also be used.
- Persons well skilled in the art will readily be able to determine what assays to use to establish activity of a compound targeting a defined disorder.
- the above stated assays should not be considered as limiting the scope of
- compositions for treating a condition characterized by oxidative stress comprise a redox-active molecule characterized by an oxidation potential, an oxidative reversibility, a reduction potential and the ability to protect energetically competent cells as described herein, said molecules being identified by the method of the present invention.
- ischemia includes but is not limited to central nervous system ischemia resulting from cardiac arrest, hypoxemia, transient Ischemic attack, stroke or severe hypotension; cerebral ischemia including stroke which may result in some degree of brain damage; ischemic heart disease (myocardial ischemia); spinal cord ischemia and paraplegia; retinal ischemia including age-related macular degeneration (ARMD); hepatic ischemia; renal ischemia; dermal ischemia; penile ischemia; pulmonary ischemia; gastric ischemia; Intestinal ischemia; splenic ischemia; pancreatic ischemia; skeletal muscle ischemia; and ischemia associated with diabetic ulcers, gangrenous conditions, post-trauma syndrome, benign prostatic hyperplasia of hypertrophy (BPH), post prostate cancer surgery, cardiac arrest resuscitation, peripheral nerve damage or neuropathies.
- ischemic heart disease myocardial ischemia
- neurodegenerative diseases or disorders refer to diseases or disorders characterized by a loss of neurons and may or may not include an inflammatory process.
- Neurodegenerative diseases or disorders include stroke, head trauma, cerebral hypoxia, spinal cord injury, epilepsy, senile dementia, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), cerebral amyloid angiopathy, HIV-related dementia, Parkinson's disease, Friedreich's ataxia or other degenerative ataxias, amyloidoses, Leber's hereditary optic neuropathy (LHON), Huntington's disease, prion diseases, myasthenia gravis, Down's syndrome, prion diseases including Creutzfeldt-Jakob disease, Tay-Sach's disease, diabetic neuropathy, neuropathic pain, encephalitis, meningitis, and Duchenne's muscular dystrophy.
- profile includes one or more parameters or measurements.
- terapéuticaally effective amount means an amount of a compound or composition effective to reduce or alleviate the symptoms of interest.
- treatment means any treatment of a syndrome, disease or disorder in a mammal, including: preventing or protecting against the syndrome, disease or disorder, that is causing the clinical symptoms of the disease to develop; inhibiting the disease, that is, arresting or suppressing the development of clinical symptoms; and/or relieving the disease, that is, causing the regression of clinical symptoms.
- a “flavonoid core structure” is a scaffold of the following structure:
- quercetin Some examples within this group are quercetin, luteolin, hesperetin, 8-acetyl quercetin, 6,8-dibromo-quercetin, anthocyanidin, and 8-(2-hydroxy)-ethyl quercetin.
- quinone core is a scaffold including a cyclohexadiene-dione moiety.
- the term “quinone core” includes but is not limited to the o-quinone scaffold, the p-quinone scaffold, the naphthoquinone scaffold and the anthraquinone scaffold.
- stilbene core is a scaffold of the following structure:
- disorder means any disease, condition, symptom, or indication.
- compositions described herein may be administered by any number of routes Including but not limited to, oral, intravenous, Intramuscular, intra-arterial, intramedullary, intrathecal, intraventricular, transdermal, subcutaneous, Intraperitoneal, Intranasal, enteral, topical, sublingual or rectal means.
- these pharmaceutical compositions may contain suitable therapeutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically, therapeutically, or neutraceutically. Further detail on technique for formulation and administration may be found in the latest edition of Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, Pa ).
- compositions described herein include compositions wherein the active Ingredients are contained in an effective amount to achieve the intended purpose.
- the determination of an effective dose is well within the capability of those skilled in the art.
- the therapeutically effective dose can be estimated initially either in cell culture assays, e.g. neuronal cells, or in animal models, usually mice or rats.
- the animal model may also be used to determine the appropriate concentration range and route of administration. Such information can then be used to determine useful doses and routes of administration.
- Administration of the compositions can be via any of the accepted modes of administration for agents that serve similar utilities.
- the therapeutically acceptable composition will contain about 0.1% to 90%, preferably about 0.5% to 50%, by weight of a redox-active molecule, the remainder being suitable pharmaceutical excipients, carriers, etc.
- a therapeutically acceptable, non-toxic composition is formed by the incorporation of any of the normally employed excipients, such as, for example, mannitol, lactose, starch, magnesium stearate, sodium saccharine, talcum, cellulose, sodium crosscarmellose, glucose, gelatin, sucrose, magnesium carbonate, and the like.
- excipients such as, for example, mannitol, lactose, starch, magnesium stearate, sodium saccharine, talcum, cellulose, sodium crosscarmellose, glucose, gelatin, sucrose, magnesium carbonate, and the like.
- Such compositions take the form of solutions, suspensions, tablets, dispersible tablets, pills, capsules, powders, sustained release formulations and the like.
- Liquid therapeutically administrable compositions can, for example, be prepared by dissolving, dispersing, etc. a redox-active compound and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline, aqueous dextrose, glycerol, glycols, ethanol, and the like, to thereby form a solution or suspension.
- a carrier such as, for example, water, saline, aqueous dextrose, glycerol, glycols, ethanol, and the like
- the pharmaceutical composition to be administered may also contain minor amounts of nontoxic auxiliary substances such as wetting agents, emulsifying agents, or solubilizing agents, pH buffering agents and the like, for example, sodium acetate, sodium citrate, cyclodextrine derivatives, sorbitan monolaurate, triethanolamine acetate, triethanolamine oleate, etc.
- Dosage forms or compositions containing active ingredient in the range of 0.005% to 95% with the balance made up from non-toxic carrier may be prepared.
- a therapeutically acceptable non-toxic composition is formed by the incorporation of any of the normally employed excipients, such as, for example pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, talcum, cellulose derivatives, sodium crosscarmellose, glucose, sucrose, magnesium carbonate, sodium saccharin, talcum and the like.
- excipients such as, for example pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, talcum, cellulose derivatives, sodium crosscarmellose, glucose, sucrose, magnesium carbonate, sodium saccharin, talcum and the like.
- Such compositions take the form of solutions, suspensions, tablets, capsules, powders, sustained release formulations and the like.
- Such compositions may contain 0.01%-95% active ingredient, preferably 0.1-50%.
- the solution or suspension in for example propylene carbonate, vegetable oils or triglycerides, is preferably encapsulated in a gelatin capsule.
- a gelatin capsule Such diester solutions, and the preparation and encapsulation thereof, are disclosed in U.S. Patents Nos. 4,328,245 ; 4,409,239 ; and 4,410,545 .
- the solution e.g. in a polyethylene glycol, may be diluted with a sufficient quantity of a therapeutically acceptable liquid carrier, e.g. water, to be easily measured for administration.
- liquid or semi-solid oral formulations may be prepared by dissolving or dispersing the redox-active compound in vegetable oils, glycols, triglycerides, propylene glycol esters (e.g. propylene carbonate) and the like, and encapsulating these solutions or suspensions in hard or soft gelatin capsule shells.
- the formulation can be administered in a single unit dosage form for continuous treatment or in a single unit dosage form ad libitum when relief of symptoms is specifically required.
- the formulation may be administered as a bolus or as a continuous intravenous infusion after onset of symptoms of stroke, myocardial infarction or chronic heart failure.
- Parenteral administration is generally characterized by injection, either subcutaneously, intramuscularly or intravenously.
- Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions.
- Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol or the like.
- the pharmaceutical compositions to be administered may also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, solubility enhancers, and the like, such as for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate, cyclodextrins, etc.
- parenteral administration employs the implantation of a slow-release or sustained-release system, such that a constant level of dosage is maintained. See, e.g., U.S. Patent No. 3,710,795 .
- the percentage of active compound contained in such parenteral compositions is highly dependent on the specific nature thereof, as well as the activity of the compound and the needs of the subject. However, percentages of active ingredient of 0.01% to 10% in solution are employable, and will be higher if the composition is a solid which will be subsequently diluted to the above percentages.
- the composition will comprise 0.2-2% of the active agent in solution. Nasal solutions of the active compound alone or in combination with other therapeutically acceptable excipients can also be administered.
- the cyclic voltammetry measurement was conducted in a three-electrode system comprising a microelectrode made from platinum (a disk of 1.6mm 2 area), a counter electrode made from a coiled platinum wire and a silver/silver chloride reference electrode, on a voltammetric analyzer Epsilon with a C-3 cell , all from Bioanalytical Systems (West Lafayette, IN).
- TBAP tetrabutyl ammonium perchlorate
- DMF N, N-dimethyl formamide
- the reference electrode was kept in a 3 M solution of NaCl in distilled water.
- the reference electrode was rinsed with water, then methanol, and dried by air blow.
- the reference electrode was gently shaken, if necessary, to remove any air bubble within the base.
- the auxiliary electrode was rinsed with water, then methanol and dried by air blow.
- the working electrode was polished on a nylon pad coated with a 1 ⁇ -meter diamond slurry in oil, then rinsed thoroughly with methanol. It is then polished on a corduroy pad coated with a 0.15 ⁇ -meter alumina slurry in water, then rinsed thoroughly with water and methanol, then dried by air blow.
- the electrochemical cell Prior to beginning a measurement and when changing compound solutions, the electrochemical cell was thoroughly cleaned with methanol, including glass reservoir, Teflon cap, and purge lines. Likewise, all electrodes were cleaned with methanol upon removal for any reason, including prior to polishing the working electrode between runs. While the working electrode was being polished, the reference electrode was removed from the test solution, rinsed with methanol and water, and stored in aqueous 3M NaCl solution.
- the solution containing the compound was transferred into the cell, where the three electrodes were immersed.
- the solution was stirred and purged with dry argon for at least 1 minute to remove any oxygen, after which a blanket of argon was kept above the solution to prevent any diffusion of oxygen. Stirring was halted prior to collecting data. Care was taken to ensure there were no bubbles attached to any of the electrodes prior to beginning electrochemical measurements.
- the solution is not a perfect conductor, and its resistance R introduces a voltage drop when a current flows between the working to the auxiliary electrode. This voltage drop introduces an error between the nominal voltage imposed by the voltammetric analyzer and the actual voltage between the working and auxiliary electrodes.
- the instrument can automatically measure and compensate for this iR drop, using the "iR Compensation" feature of the software. Normal values are the following:
- the cyclic voltammogram was recorded. After each cyclic voltammogram was recorded, the working electrode was polished again, as indicated above, to regenerate a clean, electroactive surface.
- the square wave voltammetry measurement was conducted in a three-electrode system comprising a microelectrode made from platinum (a disk of 1.6mm 2 area), a counter electrode made from coiled platinum wire and a silver/silver chloride reference electrode, on a voltammetric analyzer CV-50W with a C-3 cell , all from Bioanalytical Systems (West Lafayette, IN).
- TBAP tetrabutyl ammonium perchlorate
- DMF N,N-dimethyl formamide
- the solution is stirred and sonicated if necessary to make sure that the compound is dissolved to the best of its ability.
- the reference electrode is kept in a 3 M solution of NaCl in distilled water.
- the reference electrode is rinsed with water, then methanol, and dried by air blow.
- the reference electrode is gently shaken, if necessary, to remove any air bubble at the base.
- the auxiliary electrode is rinsed with water, then methanol and dried by air blow.
- the working electrode is polished on a nylon pad coated with a 1 ⁇ -meter diamond slurry in oil, then rinsed thoroughly with methanol. It is then polished on a corduroy pad coated with a 0.15 ⁇ -meter alumina slurry in water, then rinsed thoroughly with water and methanol, then dried by air blow.
- the electrochemical cell Prior to beginning and when changing compound solutions, the electrochemical cell should be thoroughly cleaned with methanol including the glass reservoir, Teflon cap, and purge lines. Likewise, all electrodes should be promptly cleaned with methanol upon removal for any reason, including prior to polishing the working electrode between runs. While the working electrode is being polished, the reference electrode should be removed from the test solution, rinsed with methanol and water, and stored in aqueous 3M NaCl solution.
- the solution containing the compound is transferred into the cell, where the three electrodes are immersed.
- the solution is stirred and purged with dry argon for at least 1 minute to remove any oxygen, after which a blanket of argon is kept above the solution to prevent any diffusion of oxygen. Stirring is halted prior to collecting data. Care must be taken to ensure there are no bubbles attached to any of the electrodes prior to beginning electrochemical measurements.
- the voltammogram can then be recorded. After each square wave voltammogram is recorded, the working electrode is polished again, as indicated above, to regenerate a clean, electroactive surface.
- Endothelial-Leukocyte Adhesion Molecule also known as E-selectin, is expressed on the surface of endothelial cells.
- LPS lipopolysaccharide
- IL-1 ⁇ IL-1 ⁇
- test agents were tested for their abilities to reduce this expression, in accordance with studies showing that reduction of leukocyte adhesion to endothelial cell surface was associated with decreased cellular damage (e.g., Takada, M. et al., Transplantation 64: 1520-25, 1997 ; Steinberg, J.B. et al., J. Heart Lung Trans. 13:306-313, 1994 ).
- Endothelial cells may be selected from any of a number of sources and cultured according to methods known in the art; including, for example, coronary artery endothelial cells, human brain microvascular endothelial cells (HBMEC; Hess, D.C. et al., Neurosci. Lett. 213(1): 37-40, 1996 ), or lung endothelial cells.
- HBMEC human brain microvascular endothelial cells
- HBMEC human brain microvascular endothelial cells
- IL-1 ⁇ lung endothelial cells.
- Endothelial cells may be selected from any of a number of sources and cultured according to methods known in the art; including, for example, coronary artery endothelial cells, human brain microvascular endothelial cells (HBMEC; Hess, D.C. et al., Neurosci. Lett. 213(1): 37-40, 1996 ), or lung endothelial cells.
- Cells were conveniently cultured
- Treatment buffer was removed and replaced with pre-warmed Fixing Solution® (100 ⁇ L/well) for 25 minutes at room temperature. Cells were then washed 3X, then incubated with Blocking Buffer (PBS + 2% FBS) for 25 minutes at room temperature. Blocking Buffer containing Monoclonal E-Selectin Antibody (1:750, Sigma Catalog #S-9555) was added to each well. Plates were sealed and stored at 4° overnight. Plates were washed 4X with 160 ⁇ L Blocking Buffer per well. Second Antibody-HRP diluted 1:5000 in Blocking Buffer was then added (100 ⁇ L/well), and plates were incubated at room temperature (protected from light) for two hours.
- Blocking Buffer PBS + 2% FBS
- This assay was used to induce ischemia by anoxia-reoxygenation in cultured hippocampal neuronal cells. Test compounds were added to assess potency and efficacy against ischemia-induced neuronal cell injury and cell death.
- Neurobasal/B27i Neurobasal medium (available from Invitrogen, San Diego, CA) with 1x B27 supplement (Invitrogen), 0.5 ⁇ M L-glutamine, 25 ⁇ M L-glutamic acid, and 1 x Penicillin/Streptomycin.
- HSS Hank's Basic Salt Solution
- Ca/Mg-free Hank's Basic Salt Solution
- HEPES 10 mM, pH 7.3
- sodium bicarbonate 0.5%
- 1X Penicillin/Streptomycin 1 mM MEM sodium pyruvate.
- Poly-D-lysine (Sigma, St. Louis, MO), 50 ⁇ g/ml solution.
- Plastic Culture Flasks T75 cm 2 ) or 24-well cell culture plates treated with Poly-D-Lysine (Sigma, St. Louis, MO).
- a pregnant female mouse (E18-E19) was euthanized with CO 2 followed by removal of the uterus, which was then placed in a sterile plastic petri dish.
- the embryos were removed from the sac, and the embryonic brains removed and immersed in cold (4°C) Buffered Salt Solution (HBSS; Ca/Mg free; Life Technologies) in a small petri dish.
- HBSS cold (4°C) Buffered Salt Solution
- Hippocampi were then removed from the brains under a dissecting microscope and placed on a paraffin-covered dish.
- the meninges were stripped away and the dissected hippocampi were collected in a small petri dish in HBSS.
- the hippocampi were transferred to a 15-ml centrifuge tube (normally 10-12 brains) filled with HBSS.
- the tube containing the brains was centrifuged at 1000 rpm for 2 min in a tabletop centrifuge. The supernatant was removed, 2 ml of HBSS was added to the hippocampi in the tube, and the resulting suspension was triturated 2 times each with long-tipped siliconized glass pipettes having progressively smaller apertures, starting with a pipette with a standard size opening (approximately 1.0 mm diameter), following with one having an aperture of half standard size (approximately 0.5 mm diameter), then with one having an aperture about one-half that size (0.25 mm diameter).
- the suspension was then centrifuged again at 1000 rpm for 2 min in a tabletop centrifuge, the supernatant was discarded, and 2 ml of Neurobasal/B27i (with antibiotics) was added to the tube. The trituration procedure described above was then repeated on this suspension.
- the density of cells was determined on a small aliquot of cells using standard counting procedures and correcting for cell viability by trypan blue stain exclusion. Using this procedure, the expected yield is 3 ⁇ 10 5 - 6 ⁇ 10 5 cells/brain.
- Cells were then added to PDL-coated 24 well plates, flasks or MetTek dishes in Neurobasal/B271 at a density of about 1.5 ⁇ 10 6 cells (T75 flask) or about 100,000 cells/well of a 24-well plate. Plated cells were incubated at 37°C in an atmosphere of 5 % CO 2 / 95 % O 2 .
- Neurobasal media NoG neurobasal media, B27 supplement and B27 Supplement minus AO (Invitrogen).
- Neurobasal/B27 medium is prepared with 2X B27 minus AO supplement, 0.5 mM L-glutamine and 0.25X penicillin/streptomycin.
- Cell Tracker Green was obtained from Molecular Probes and a fresh 5 ⁇ M solution was prepared from 10 mM stock just before use.
- NoG-Neurobasal contains NoG neurobasal medium plus 0.5 mM glucose, 0.1 mM L-glutamine and 0.25X Penicillin/Streptomycin.
- Primary hippocampal neuronal cells were prepared according to the methods described above and were cultured in poly-D-lysine coated 24 well plates for 10-11 days prior to use.
- LoG-Neurobasal medium 100 ml was prepared by pre-equilibrating the medium in a T150 cm 2 flask in a hypoxic chamber overnight. Following pre-incubation under hypoxic conditions, the LoG-Neurobasal media was lightly bubbled with 100% N 2 for 30 min to completely deoxygenate the media. An additional 20 ml LoG-Neurobasal was pre-equilibrated in a T75 cm 2 flask and 100 ml Neurobasal/B27AO was incubated in a normal incubator (5% CO 2 overnight. Reoxygenated medium was prepared by placing medium overnight in the culture incubator (5% CO 2 /95% O 2 ) prior to use.
- normoxia controls pre-equilibrated normoxic LoG-Neurobasal medium was added to each well of cells, and the plate was replaced in the normal culture incubator for 5 hr. After 5 hr of hypoxia, the existing media was carefully aspirated off, and 2mL of new, re-oxygenated (pre-equilibrated) Neurobasal/B27AO was added to each well. The same test compounds (in the same the concentrations) were added back into the corresponding wells. Plates were placed in the cell culture incubator (5% CO 2 /95% O 2 ) and reoxygenated for 20-24 hr. After reoxygenation for 20-24 hr, live neurons were quantitated using the cell tracker green fluorescence method, described below.
- HBSS pH 7.4, pre-warmed to 30-37°C
- 5 ⁇ M Cell Tracker Green fluorescent dye dissolved in HBSS. Plates were placed in the dark at room temperature for 15 minutes, and are then washed with two milliliters of HBSS. One milliliter of HBSS was then added to each well, and fluorescent cells were counted using a fluorescent microscope. Significantly increased cell viability compared to control cells was indicative of a protective compound.
- HGOS High Glutamate-Induced Oxidative Stress Assay
- HGOS high glutamate-induced oxidative stress
- the cells were seeded into 96-well plates at a density of 2000 per well and left to grow for 72 hours in a 33°C incubator with 5% CO 2 in air atmosphere.
- the passage number of the cells for each assay experiment were no later than p11 in order to minimize experimental variation.
- VWRBrand DyNA Block 1000 deep well mother plates (VWR Cat. # 40002-008) were used for the preparation of the test compounds.
- DMEM-No Glu containing 1mM glucose, 30 mM glutamate and 1x Pen/Strep.
- DMEM-No Glu with 1mM glucose and 1x P/S was used as the negative control
- DMEM-No Glucose with 1 mM glucose, 100 M glutamate was used as a positive control and 100 ⁇ M Glutathione was added to the positive control as a standard. All of the procedures for this involving the making and dilution of compounds were performed using aseptic conditions and with minimal light.
- the plates were removed from the incubator and examined under the microscope for morphological appearance and density. Using an aseptic technique and an 8-channel aspirator the media was carefully removed from the cells and replaced with 200 ⁇ l of 1x HBSS. This was done as quickly as possible to prevent the cells drying out. The plates were then placed in the humidified 37°C incubators of the Biomek 2000 Side Loader. Four plates were washed at a time so as to minimize the time that the cells were sitting in 1x HBSS prior to addition of the compound test solution.
- the Beckman Biomek workstations were used to load the compounds and controls from the mother plates onto the cell plates that were prewashed with HBSS under sterile conditions.
- the plates were incubated in the upper HTS incubator at 37°C in 5% CO 2 for exactly 16 hrs. The following day, using the Beckman Biomek workstations, the plates were removed from the incubator. Using Cell Tracker Addition, the compounds were removed from the plates, washed once with 200 ⁇ M of pre-warmed 1x HBSS and then 100 ⁇ L of 5 ⁇ M Cell Tracker Green was added to each well.
- the plates were incubated at 37°C for 30 min to allow the dye to enter the cell and be cleaved by the esterases. After washing the cells twice with prewarmed 1x HBSS, the plates were read with the 485 excitation; 538 emission filter pair on a Fluoroskan.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- General Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Neurology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Neurosurgery (AREA)
- Molecular Biology (AREA)
- Dermatology (AREA)
- Physical Education & Sports Medicine (AREA)
- General Physics & Mathematics (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Diabetes (AREA)
- Rheumatology (AREA)
- Cardiology (AREA)
- Pulmonology (AREA)
- Psychology (AREA)
Description
- The present invention is directed to novel methods for Identifying therapeutic compounds based on their structural and physlcal-chemical properties.
- In response to the ever increasing demand for novel compounds useful in the effective treatment of various disorders, a variety of strategies for discovering and optimizing new therapeutics has been developed. For the most part these strategies are dependent upon techniques that allow identification of molecules binding to a given biological target.
- In one such strategy, novel drugs are Identified by screening compound libraries and determining which compounds have the highest therapeutic properties, and optimizing such properties by synthesizing structurally related analogs. The limitation of such an approach is that it is possible to synthesize and test only a very small subset of all possible molecules thereby resulting in a high probability that the most efficacious molecules will be missed.
- In another approach novel drugs are Identified by determining structure activity relationships (SAR) correlating a common structural feature of the molecule to target based biological activity. While widely used this method does not always yield active analogs indicating SAR alone may not be sufficient to warrant biological activity In all cases.
- SAR activity by NMR as described In
US 5,698,401 , relates to a process for Identifying compounds which bind to a specific target molecule. In this approach the physical structure of a target protein is determined by nuclear magnetic resonance analysis (NMR) and the small molecule building blocks are Identified that bind to the protein at nearby points on the protein surface. Adjacently binding small molecules are then coupled together with a linker in order to obtain compounds that bind to the target protein with higher affinity that the unlinked compounds alone. Thus, by having available the NMR structure of the target protein, the lengths of linkers for coupling two adjacently binding molecules can be determined and small molecules can be rationally designed. Although these methods are powerful they have serious limitations, such as the required amounts of target protein and the fact that the protein must be 15N-labeled to be useful for NMR studies. - With the advent of combinatorial chemistry and high throughput screening, numerous high-profile reviews have appeared In the literature that classify and prioritize the chemical space for drug discovery. Lipinski et al., Adv. Drug. Deliv. Rev., 3-25, 1997, Is a widely cited review that correlates physical-chemical properties of permeability and absorption with biological activity. This review defines the "rule of 5" which a molecule should generally not violate to be granted the status of "drug-like" molecule. The "rule of 5" states that poor absorption or permeation are more likely when there are more than 5-H bond donors (expressed as the sum of OHs and NHs); more than 10 H-bond acceptors (expressed as the sum of Ns and Os); the molecular weight is over 500; and the log P is greater than 5.
- Ghose et al., J. Comb. Chem., 1, 55-68, 1999, provides an analysis of some computable physical-chemical properties and chemical constitutions of known drug molecules available in the comprehensive Medicinal Chemistry (CMC) database and seven known drug classes. Their study showed qualifying ranges for calculated log P, refractivity, molecular weight, and total number of atoms.
- In recent years, electronically active molecules, such as antioxidants and reductants, have been recognized as functioning In redox regulation of key biological processes such as Immune response, cell-cell adhesion (e.g. atherosclerosis), cell proliferation, inflammation, metabolism, glucose uptake (diabetes), and programmed cell death (apoptosis).
- It has been described in the art, that biological activity of certain compounds is related to their capacity to accept one or two electrons to form the corresponding radical anion or dianion, and that the electron-accepting capacity of these substances can be modified by directly adding substituents to the core structure. For these types of compounds, the attracting or donor effects of the substituents are very Important In affecting the redox properties of the core structure system, either facilitating or Interfering with the electron transfer, (see for example a study of the substituent effect on the redox properties of the quinone moiety, Aguilar-Martinez et al., J.Org.Chem., 64, 3684-3694, 1999). If a molecule's core structure is affected by its substituents, a change of its voltammogram may occur representing a change in electron transfer (redox) properties.
- The redox behavior of a series of structurally related flavonoids under physiological conditions has been investigated by Hodnik, W.F. et al, Biochem. Pharmacol., 37 (13), 2607-11, 1988, as well as the relationship of flavonoids oxidation potential and effect on the hepatic metabolism, see Hendrickson, H.P. et al., J. Pharm. Biomed. Anal., 12(3), 335-41, 1994, Half peak oxidation of flavonoids was measured and correlated to LPO inhibition data in Van Acker et al., Free Radic. Biol. Med., 20 (3), 331-342, 1996 and the redox Intermediates of flavonoids and caffeic acid esters from propolis were studied by cyclic voltammetry, see Rapta, P. et al., Free Radic. Biol. Med., 18(5), 901-8, 1995.
- Structural activity relationship studies on apomorphine and its derivatives have been described by Lashuel, H.A. et al., Journal of Biol. Chem., 277(45), 42881-42890, 2002.
- Ashnagar A. et al., Biochim. Biophys. Acta, 801, 351-9, 1984, have described the measurements of reduction potentials of hydroxy-1,4-naphthoquinones and hydroxy-9,10-anthraquinones as well as the corresponding methoxy- and acetoxyquinones, and the role of internal hydrogen bonding and its bearing on the redox chemistry of the anthracycline antitumor quinones. The correlation among antitumor activity, quinone reduction potential and the logarithm of the partition coefficient (log P) was obtained by Kunz et al., J. Med. Chem., 34, 2281-6, 1991. The relationships of reductive potential, kinetics of enzymatic reduction, augmented oxygen consumption and cytotoxicity were determined for seven clinically relevant mitomycin antibiotics by Pan S.S. et a/., Mol. Pharmaco/., 37, 966-70,1997. Twelve 1,4-naphthoquinones were tested against the ascetic form of sarcoma and it was shown by statistical analysis that the most important molecular parameterw determining their effectiveness in prolonging the life of mice bearing this tumor were their redox potentials, see Hodnett E.M. et al. J. Med. Chem., 26, 570-4, 1983. Electrochemical properties of some biologically active quinone derivatives, furanquinones, pyridoquinones and diplamine, a cytotoxic pyridoacridine alkaloid, have been reported in Crawford, P.W. et al., J. Electrochem. Soc., 144, 3710-3715, 1997, indicating a possible relationship between reduction potential and anticancer activity.
- Cyclic voltammetry has been used for the detection of compounds In different solutions, see for example, Kilmartin, P. Antioxidants and Redox Signaling, 3, (6), 941-955, 2001, and in a redox control and monitoring high throughput screening platform used In conjunction with another detector, (see
US Application 2002/0123069 ), but heretofore cyclic voltammetry has not been used for the a priori identification of possible therapeutic candidates. - It is evident that there is a need for novel techniques useful for rapidly and efficiently identifying molecule compounds that are capable of having a therapeutic effect.
- It has been surprisingly found that novel therapeutic molecules can be identified by their physical-chemical properties comprising a least one redox parameter and failing within a range predefined by the physical-chemical/biological relationship of a previously tested small subset of compounds with the same core structure.
- The present Invention relates to a rapid and efficient method of Identifying therapeutic compounds, by allowing only the most favorable molecules initially selected based on their core structure and their physical-chemical characteristics to be further assayed.
- Accordingly, the present invention provides a method of identifying and selecting therapeutic compounds having a predetermined core structure, said method comprising:
- establishing a relationship between physical-chemical profile and biological activity; wherein the physical-chemical profile comprises one or more parameters selected from onset of oxidation, potential of oxidation, potential of reduction, reversibility of oxidation, reversibility of reduction, current of oxidation or current of reduction; and wherein the biological activity is measured in an assay effective in detecting compounds for the treatment of a targeted disorder;
- testing further potential therapeutic candidates with said core structure for their physical-chemical properties; and
- selecting therapeutic compounds based on their physical-chemical parameters falling within a range predefined by the physical-chemical/biological relationship of the previously tested subset of compounds,
- wherein the predetermined core structure is selected from a quinone core structure, the targeted disorder is oxidative stress, inflammation, ischemia, neurodegeneration, or Alzheimer's disease, and the physical-chemical profile of the compounds comprises a parameter for the total reversibility of reduction of 75% or more, said parameter for reversibility of reduction being a ratio of reduction wave maximum current to oxidation wave maximum current of 0.75 or more;
- or wherein the predetermined core structure is selected from a core structure of Formula I:
- wherein the targeted disorder is oxidative stress, inflammation, neurodegeneration, or ischemia, and wherein
(I-a) additional substitution at the phenyl rings of the core structure of Formula I does not include a nitro group; and the physical-chemical profile of the compounds comprises the parameter for potential of the first oxidation wave where said potential falls below 1000mV versus a silver/silver chloride reference electrode; or
(I-b) additional substitution at the phenyl rings of the core structure of Formula I includes a nitro group and the physical-chemical profile of the compounds comprises one or more parameters selected from the parameter for potential of the first oxidation wave where said potential falls between 950mV and 1250mV versus a silver/silver chloride reference electrode and the parameter for reversibility of the first oxidation wave measuring more than 20%, said parameter for reversibility of the first oxidation wave being a ratio of first oxidation wave maximum current to reduction wave maximum current of 0.2 or more;
or wherein the predetermined core structure is selected from a core structure of Formula III:
(III-b) the physical-chemical profile of the compounds comprises the parameter for potential of the first reduction where said potential is more negative than -790mV versus a silver/silver chloride reference electrode
or wherein the predetermined core structure is selected from a core structure of Formula IV:
(IV-a) the physical-chemical profile of the compounds comprises the parameter for oxidation potential where said potential falls between 850mV and 1200my versus a silver/silver chloride reference electrode. - The features, aspects and advantages of the present invention will become better understood with regard to the following description, claims, and accompanying drawings.
-
-
FIG. 1 illustrates a cyclic voltammogram of a stilbene analog showing potential of oxidation wave, reversibility of reduction wave and potential of reduction wave. -
FIG. 2 Illustrates the graph of oxidation potential plotted against activity In log (1/EC50) In the neuronal oxidative stress assay (HGOS) of certain stilbene analogs, wherein one of the rings is further substituted with a nitro group. Certain compounds with a potential of the first oxidation wave ranging between 950 and 1250 mV show activity in the neuronal oxidative stress assay. -
FIG. 3 illustrates the graph of % oxidative reversibility plotted against activity In log (1/EC50) in the neuronal oxidative stress assay (HGOS) of certain stilbene analogs, wherein one of the rings is further substituted with a nitro group. Certain compounds with reversibility of the first oxidation wave of more than 20% show activity In the neuronal oxidative stress assay. -
FIG. 4 Illustrates the graph of oxidation potential plotted against activity in log (1/EC60) in the neuronal oxidative stress assay (HGOS) of certain stilbene analogs, wherein none of the rings is further substituted with a nitro group. Certain compounds with a potential of the first oxidation wave ranging below 1000 mV show activity In the neuronal redox assay. -
FIG. 5 Illustrates the graph of oxidation potential plotted against activity In log (I/EC60) In the neuronal oxidative stress assay (HGOS) of certain analogs with a flavonoid core structure. -
FIG 6 illustrates the graph of onset oxidation plotted against activity In EC50 in the ELAM assay of certain analogs with a flavonoid core structure wherein some of the substituents are hydroxy groups. Certain compounds with an onset of oxidation wave ranging between 350mV and 650mV show activity at EC50 under 30 µM In the ELAM assay. -
FIG 7 Illustrates the graph of onset oxidation plotted against activity in EC50 In the ELAM assay of certain compounds with a flavonoid core structure. Active flavonoids compounds wherein some of the substituents are hydroxy groups fall within an onset of oxidation range between 350mV and 650mV, and compounds wherein none of the substituents are hydroxy groups fall within an onset of oxidation range between 850mV and 1050mV. -
FIG. 8 illustrates the voltammograms of two compounds with an apomorphine core structure: apomorphine which reduces fibril growth by 99% and apocodeine which reduces the fibril growth by 5%. These voltammograms show that a minor variation in structure induces a variation in redox current. -
FIG. 9 illustrates the percent of fibril formation inhibition based on quantitative Thioflavin T binding assay in the presence of the six compounds with an apomorphine core structure: norapomorphine, 2,10,11-trihiydroxyaporphine, propylnorapomorphine, apocodeine, isocorydine, and bulbocapnine as described in Lashuel, H.A. et al., J.Bio.Chem., 227 (45), 42881-42890, 2002. -
FIG. 10 illustrates the graph of percent of fibril formation at two different concentrations (50mM and 100mM) vs. potential of first oxidation wave of seven compounds with an apomorphine core structure. The compounds with a potential of first oxidation wave under 1250mV show strong reduction of fibril formation in the Thioflavin T binding assay. -
FIG. 11 illustrates the graph of percent of fibril formation at two different concentrations (50mM and 100mM) vs. potential of first reduction wave of six compounds with an apomorphine core structure. The compounds with a potential of first reduction wave more negative than - 790mV show strong reduction of fibril formation in the Thioflavin T binding assay. -
FIG. 12 illustrates the graph of reversibility of reduction waves vs. biological activity in the ELAM assay in EC50 of compounds with a quinone core structure, wherein the total reversibility of reduction is the ratio of the current peak of reduction wave over the current peak of reoxidation waves or, in the case of more than one wave, the ratio of the mathematical addition of the current peaks of reduction waves over the mathematical addition of the current peaks of reoxidation waves. -
FIG. 13 illustrates the graph of oxidation potential plotted against activity in log (1/EC50) in the neuronal oxidative stress assay (HGOS) of certain chromans. Certain compounds with a potential of the first oxidation wave ranging between 850 and 1200 mV show activity in the neuronal oxidative stress assay. - The interaction of a molecule with a biological target may not be limited to structural recognition alone. Other types of interaction may occur, and some of them may involve the exchange of one or more charges. Because of their distinct electronic distribution, molecules exhibit different physical-chemical profiles, particularly redox profiles, i.e. the measure of an ability to give or accept charges, the number of charges exchanged, the kinetics of the process, and the subsequent molecular mechanisms that follow the addition or subtraction of charges. Various electroanalytical techniques can be employed to determine the physical-chemical profile. Cyclic voltammetry is a technique that yields quantitative information on the above parameters. The redox profile of a molecule is well characterized by its cyclic voltammogram.
- Other specific electroanalytical techniques may also be used to measure redox profile, such as for example single cyclic voltammetry, continuous cyclic voltammetry (with and without the integration of current), square wave voltammetry, square wave stripping voltammetry, AC voltammetry, choromoamperometry, chronocoulometry, chronopotentiometry, as well as various potentiostatic and galvanostatic techniques can be employed. A description of various electroanalytical techniques can be found in numerous textbooks, such as. Monk S, Fundamentals of Electroanalytical Chemistry, Wiley & Sons, New York,. 156-175, 2001 or Bard A.J. et al, Electrochemical Methods, Wiley & Sons, New York, Ch. 6, , 1980. The above stated electroanalytical techniques should not be considered as limiting the scope of the invention, but merely as being illustrative and representative thereof.
- The redox properties of a molecule can be acquired from a variety of sources including cyclic voltammetry, in which the compound is characterized by the current-potential relationship exhibited at an electrode such as for example platinum, gold, or glassy carbon electrodes. In this technique the potential of a stationary electrode is changed linearly with time, starting from a fixed potential and moving to potentials more reductive or more oxidative. After traversing the potential region in which one or more electrode reactions take place, the direction of the linear sweep is reversed and the electrode reactions of intermediates and products that may have been formed during the forward scan can be detected. This has the advantage that the product of the electron transfer reaction that occurred in the previous scan can be probed again in the current scan. Cyclic voltammetry is a simple and direct method for measuring the formal potential of a half reaction when both oxidized and reduced forms are stable during the time required to obtain the current-potential curve. The resulting plot of current versus potential is termed a cyclic voltammogram. A cyclic voltammogram is a complicated, time-dependent function of a large number of physical and chemical parameters. It will give information on the anodic and cathodic peak potentials, the half-peak potential of the oxidation wave, the peak separation, the potential midway between the anodic and the cathodic peak. Only in the case where a redox system remains in equilibrium throughout the potential scan is the voltammogram reversible. The shape of a voltammogram can be significantly altered if there are coupled chemical reactions either before or after the electrochemical process. The mechanism, the rate, and the equilibrium constants of the process can all play a part in the final shape of the voltammogram that characterizes a certain molecule. Cyclic voltammetry is a powerful tool for the determination of formal redox potentials, detection of chemical reactions that precede or follow the electrochemical reaction, and evaluation of electron transfer kinetics.
- A cyclic voltammogram yields data on the ability of the molecule to shed one or more electrons measured by the oxidation potential, the relative number of electrons exchanged measured by the oxidation current, the reversibility of the oxidation process measured by the ratio of the oxidation current to the corresponding reduction current, and the kinetics of the oxidized species measured through variation of the scan rate; as well as the ability of the molecule to accept one or more electrons measured by the reduction potential, the number of electrons exchanged measured by the reduction current, the reversibility of the reduction process measured by the ratio of the reduction current to the corresponding oxidation current; and the stability of the reduced species measured through variation of the scan rate.
- Square wave voltammetry is a technique that yields quantitative information on the ability of the molecule to shed or accept one or more electrons with a very low detection limit due to its pulsed voltammetric technique. The primary advantage of the pulse voltammetric techniques is their ability to discriminate against charging current. As a result the pulse techniques are more sensitive to oxidation and reduction currents than conventional voltammetry. Square wave voltammetry yields peaks for faradic processes, where the peak height is directly proportional to the concentration of the species In solution. This results in improved resolution for multiple analyte systems and more convenient quantization. See for example, O'Cea, J. et al "Theory of Square Wave Voltammetry for Kinetic Systems", Anal. Chem. 53(4), 695, 1981 and Krause, J et al. "Analytical Application of Square Wave Voltammetry", Anal. Chem. 41(11), 1365, 1969.
- Even greater sensitivity when conducting measurements can be attained by using Stripping Square Wave Voltammetry, in which the species of interest is concentrated into the working electrode by electrochemical means before doing the analysis. With a sufficiently long concentration step, the concentration of the substance will be much higher In the electrode than in the sample solution. If the electrode potential is then scanned, the substance will be stripped from the electrode causing an increase in the cell current as this process occurs. The advantages of square wave stripping voltammetry over linear sweep are:
- it incorporates a pulsed waveform thus enhancing the sensitivity by repeated oxidation and reduction of the same analyte species;
- it is a purely subtractive technique, thus limiting currents due to dissolved species such as oxygen not Interfere with the analytical signal;
- it is a fast technique and can be obtained in a matter of seconds;
- it provides kinetic Information because of the ability to analyze both the forward and reverse currents as well as the net current, information about reaction reversibility, and electrode structure can be obtained easily.
- In order to measure signals it is beneficial to utilize electrodes at which the competing redox reactions, such as, for example, the hydrogen evolution reaction or oxygen evolution reaction, do not interfere. This can be achieved by employing electrodes characterized by sufficiently high overpotential towards the competing reactions, such as but not limited to platinum, gold and glassy carbon. The working electrode may be stationary or rotating, of any geometry, and can Include convective mixing. The measurements can be done in a variety of systems including but not limited to oxygenated or non-oxygenated environments, and protonated and non-protonated environments.
- The electro-chemical measurement effected herein is typically conducted in a three-electrode system (working electrode/counter electrode/reference electrode) or, possibly a two electrode system (working electrode/counter electrode). The area of the counter electrode may typically be larger than that of the measuring electrode.
- The redox profiles of the compounds described herein were measured by cyclic voltammetry, as described in Example 1 or by square wave voltammetry, as described in Example 2. All stated potential values are stated versus a silver/silver chloride reference electrode described in Examples 1 and 2. Those skilled in the art will appreciate that the silver/silver chloride electrode may be substituted with other reference electrodes, and that such substitution can result in different values, but this does not depart from the true spirit and scope of the invention.
- The physical-chemical profile of this invention comprises a redox profile and its related physical measurements, optionally in conjunction with an energy profile and/or a transport profile and their related physical measurements.
- The energy profile of a molecule can be acquired from its optical values such as linear emission, linear absorption and non-linear absorption. Linear emission is measured by the wavelength of maximum fluorescence λem, i.e. the radiating energy released by the stabilization of an electron from a high energy level (LUMO) to a lower energy level (HOMO); and/or by quantum yield of fluorescence Φ, i.e. the probability of the molecule to stabilize an electron from its high energy level (LUMO) to a lower energy level (HOMO). Linear absorption is measured by the wavelength of maximum absorption λmax, i.e. the amount of energy needed to eject an electron from the main energy level of the molecule to the next higher energy level with the highest probability; and/or molar extinction coefficient ε, i.e. the probability of the molecule to eject an electron from its main energy level to next higher energy level when it receives the proper amount of energy; and/or the longest wavelength of absorption λend, i.e. the minimum energy needed to eject an electron form the main energy level of the molecule. Polarizability Δλpol is measured by the ability of the electron cloud of the molecule to be shaped by an external electromagnetic field. Polarizability measurements are conducted by solvatochromism, measuring the shift in wavelength of maximum absorption as a function of solvent polarity (e.g. between octanol and DMSO).
- The transport profile of a molecule can be acquired from its partition coefficient, and/or diffusion constant, and/or molecular weight, and/or melting point.
- In order to establish a relationship between physical-chemical profile and biological activity of a small subset of molecules, all the quantitative physical parameters of such molecules are plotted versus a particular biological assay value, and for a well defined population correlations may emerge if any of the physical parameters is related to a biological activity. The plotting may be done manually or with the help of computer programs well known in the art such as Microsoft Excel ®.
- The biological activity of the small subset of compounds can be assessed by any assay deemed relevant to the target of interest. In the present invention the biological activity of some of these compounds was determined using an in vitro model well recognized in the art for determining the degree of dysfunction of the cells when exposed to stress. In vitro models of ischemia approximate oxygen and glucose deprivation that mimic in vivo conditions, for example, by placing neuronal cultures into large anaerobic of hypoxic chambers and exchanging culture medium with de-oxygenated and defined ionic composition media. The toxic overstimulation of neuronal glutamate receptors, especially N-methyl-D-aspartate (NMDA) receptors contribute to hypoxic-ischemic neuronal injury (Choi, D.M., Neuron,1: 623-634, 1988), ischemic induction of reactive oxygen species (ROS) (Watson, B.D.et al., Ann NY Acad Sci., 59: 269-281, 1988), excessive calcium influx (Grotta, J.C., Stroke, 19: 447-454, 1988), arachidonic acid increase (Siesjo, B.K., J. Cereb. Blood Flow Metab., 1: 155-186,1981) and DNA damage (MacManus, J.P. et al., Neurosci. Lett., ,164: 89-92, 1993), each causing a cascade of neurodegeneration.
- Oxidative stress has emerged as one of the major factors in the neurodegenerative disease and may contribute to neuronal damage from ischemia (see e.g., Coyle J. T. et al., Science 262, 689 -695, 1993; or Beal M. F, Curr. Opin. Neurobiol.; 6, 661-666, 1996). Reactive oxygen species (ROS), which are generated as by-products of many metabolic processes including the mitochondrial electron transport chain, (Tan S. et al., J. Cell Biol. 141, 1423 -1432, 1998), monoamine metabolism (Maher P. et al., J. Neurosci. 16, 6394 -6401, 1996), and arachidonic acid oxidation (U Y., et al., Neuron 19,453 -463, 1997) may be the principal mediators for cell death in oxidatively stressed neuronal cells (Chan P. H., Role of oxidants In ischemic brain damage; Stroke 27,1124 -1129, 1996 and Hosler B. A. et al.; Curr. Opin. Neurol. 9, 486 -491. 1996). The damage on cellular organelles and macromolecules by chemical reactions with ROS can initiate an apoptotic program of cell death (Hosier B.A. et al., see supra) or lead to neurosis (Choi D. W. Curr: Opin. Neurobiol. 8, 667-672,1996).
- Mouse dopaminergic neuronal cell lines are useful for examining high glutarnate-Induced oxidative stress (HGOS). The cytotoxic effect of glutamate is not due to excitotoxicity, as this cell line is devoid of inotropic glutamate receptors. Rather, the glutamate-induced toxicity of dopaminergic cells is associated with an inhibition of cystine transport (Murphy T. H., et al., ), which subsequently leads to depletion of intracellular glutathione (GSH) levels (Murphy T. H., et al. ), activation of neuronal 12-lipoxygenase (U, Y. et al., see supra), Increased ROS production (Tan S. et al., see supra) and elevated intracellular Ca2+ (Li, Y. et al., see supra). Some molecules were measured for their ability to protect such cells against glutamate-induced stress and the assay is detailed In Example 5.
- Primary embryonic hippocampal neuronal cells are widely recognized as useful In models of neuronal function. The hippocampus is a source of a relatively homogenous population of neurons with well-characterized properties typical of central nervous system (CNS) neurons in general. Pyramidal neurons, the principal cell type In the hippocampus, have been estimated to account for 85% to 90% of the total neuronal population (Banker and Goslin, Culturing Nerve Cells, 2nd edition, 1998. The MIT Press, Cambridge, Massachusetts). The hippocampus also exhibits a remarkable capacity for activity-dependent changes in synaptic function, such as longterm potentiation (Hawkins RD, Kandel ER, Slegelbaum SA. Learning to modulate transmitter release: themes and variations In synaptic plasticity [review], Ann. Rev Neurosci., 16, 625-665, 1993.).
- Primary cultures of hippocampal neurons measuring the ability to protect energetically competent cells were used to test compounds for activity in neuronal protection. Some molecules were measured for their ability to protect cells against one or more standard stressors, including hypoxia and the assay is detailed in Example 4.
- For the purpose of the present invention other well known In the art in vitro cell-based assays such as inflammation assays, for example e-selectin (also named Endothelial Leukocyte Adhesion Molecule or ELAM) or C-reactive protein (CRP), or myocyte calcium-contractility assay or in vivo assays such as the rat middle cerebral artery occlusion (MCAO) model of cerebral ischemia assay, the rat paw edema assay or the mouse ear inflammation response to topical arachidonic acid assay or a skin protection assay may also be used. Persons well skilled in the art will readily be able to determine what assays to use to establish activity of a compound targeting a defined disorder. The above stated assays should not be considered as limiting the scope of the invention, but merely as being illustrative and representative thereof.
- Described herein are compositions for treating a condition characterized by oxidative stress, wherein such compositions comprise a redox-active molecule characterized by an oxidation potential, an oxidative reversibility, a reduction potential and the ability to protect energetically competent cells as described herein, said molecules being identified by the method of the present invention. Diseases, disorders, or syndromes associated with oxidative stress include, but are not limited to reperfusion injury following ischemia, myocarditis, cardiomyopathy, acute endocarditis, pericarditis, congestive heart failure, inflammatory complications of diabetes mellitus, amyetrophic lateral sclerosis, neurodegenerative diseases, such as Alzheimer's disease and dementia, autoimmune disease, Sjogren's syndrome, retinal oxidative damage, retinopathy, Crohn's disease, ulcerative colitis, angiogenesis, disorders of the peritoneal, pelvic and pleural cavity, adult respiratory distress syndrome (ARDS), lung disorders, bronchial hyperreactivity, chronic obstructive pulmonary disease (COPD), and inflammatory conditions as described herein.
- As used herein, "inflammation" or "Inflammation conditions" includes but Is not limited to muscle fatigue, autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, osteoarthritis, inflammatory bowel disease, autoimmune diabetes, skin inflammation, such as atopic dermatitis, contact dermatitis, allergic dermatitis, xerosis, eczema, rosacea, seborrhea, psoriasis, thermal and radiation burns, acne, oily skin, wrinkles, excessive cellulite, excessive pore size, intrinsic skin aging, photo aging, photo damage, harmful UV damage, keratinization abnormalities, alopecia, dyspigmentation, inflammation due to wounds, scarring or stretch marks, loss of elasticity, skin atrophy and gingivitis.
- As used herein, "ischemia" includes but is not limited to central nervous system ischemia resulting from cardiac arrest, hypoxemia, transient Ischemic attack, stroke or severe hypotension; cerebral ischemia including stroke which may result in some degree of brain damage; ischemic heart disease (myocardial ischemia); spinal cord ischemia and paraplegia; retinal ischemia including age-related macular degeneration (ARMD); hepatic ischemia; renal ischemia; dermal ischemia; penile ischemia; pulmonary ischemia; gastric ischemia; Intestinal ischemia; splenic ischemia; pancreatic ischemia; skeletal muscle ischemia; and ischemia associated with diabetic ulcers, gangrenous conditions, post-trauma syndrome, benign prostatic hyperplasia of hypertrophy (BPH), post prostate cancer surgery, cardiac arrest resuscitation, peripheral nerve damage or neuropathies.
- As used herein the terms "neurodegenerative diseases or disorders, or neurodegeneration" refer to diseases or disorders characterized by a loss of neurons and may or may not include an inflammatory process. Neurodegenerative diseases or disorders include stroke, head trauma, cerebral hypoxia, spinal cord injury, epilepsy, senile dementia, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), cerebral amyloid angiopathy, HIV-related dementia, Parkinson's disease, Friedreich's ataxia or other degenerative ataxias, amyloidoses, Leber's hereditary optic neuropathy (LHON), Huntington's disease, prion diseases, myasthenia gravis, Down's syndrome, prion diseases including Creutzfeldt-Jakob disease, Tay-Sach's disease, diabetic neuropathy, neuropathic pain, encephalitis, meningitis, and Duchenne's muscular dystrophy.
- As used herein, "physical-chemical profile" includes but is not limited to redox profile comprising one or more parameters selected from onset of oxidation, potential of oxidation or reduction waves, reversibility of said oxidation or reduction waves, and current of said oxidation or reduction waves, optionally in conjunction with energy profile and/or transport profile, as defined herein.
- As used herein, "onset of oxidation" means the potential at which the current is 1% of the maximum current.
- As used herein, "profile" includes one or more parameters or measurements.
- As used herein "redox parameter" or "redox property" or "redox" means a quantity related to a redox process that can be measured, e.g. potential, current, reversibility.
- As used herein, "reversibility" means the measure of the reaction kinetics. It may be monitored by one or more of the following parameters, ratio of the peak currents at the anodic peak and cathodic peaks, half-width of the peaks, and/or separation of the peaks. The "reversibility of oxidation wave" as described herein, is measured by the ratio of current peak of the oxidation wave to the corresponding current peak of the reduction wave, or, in the case of more than one wave, by the ratio of the mathematical addition of the current peaks of reduction waves over the mathematical addition of the current peaks of reoxidation waves.
- As used herein, "therapeutically effective amount" means an amount of a compound or composition effective to reduce or alleviate the symptoms of interest.
- As used herein, "treatment" or "treating" means any treatment of a syndrome, disease or disorder in a mammal, including: preventing or protecting against the syndrome, disease or disorder, that is causing the clinical symptoms of the disease to develop; inhibiting the disease, that is, arresting or suppressing the development of clinical symptoms; and/or relieving the disease, that is, causing the regression of clinical symptoms.
-
- Some examples within this group are quercetin, luteolin, hesperetin, 8-acetyl quercetin, 6,8-dibromo-quercetin, anthocyanidin, and 8-(2-hydroxy)-ethyl quercetin.
- As used herein a "quinone core" is a scaffold including a cyclohexadiene-dione moiety. The term "quinone core" includes but is not limited to the o-quinone scaffold, the p-quinone scaffold, the naphthoquinone scaffold and the anthraquinone scaffold.
-
- The term "stilbene core" includes cis and trans (or Z and E) single isomers, as well as a mixture of isomers.
- As used herein, "disorder" means any disease, condition, symptom, or indication.
- The pharmaceutical compositions described herein may be administered by any number of routes Including but not limited to, oral, intravenous, Intramuscular, intra-arterial, intramedullary, intrathecal, intraventricular, transdermal, subcutaneous, Intraperitoneal, Intranasal, enteral, topical, sublingual or rectal means. In addition to the active ingredients, these pharmaceutical compositions may contain suitable therapeutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically, therapeutically, or neutraceutically. Further detail on technique for formulation and administration may be found in the latest edition of Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, Pa).
- Pharmaceutical compositions described herein include compositions wherein the active Ingredients are contained in an effective amount to achieve the intended purpose. The determination of an effective dose is well within the capability of those skilled in the art. For any compound, the therapeutically effective dose can be estimated initially either in cell culture assays, e.g. neuronal cells, or in animal models, usually mice or rats. The animal model may also be used to determine the appropriate concentration range and route of administration. Such information can then be used to determine useful doses and routes of administration.
- The exact dosage will be determined by the practitioner in light of factors related to the subject that requires treatment. Dosage and administration are adjusted to provide sufficient levels of the active moiety or to maintain the desired effect. Factors which may be taken into account include the severity of the disease state, general health or the subject, age, weight, and gender of the subject, diet, time and frequency of administration, drug combination(s), reaction sensitivities, and tolerance/response to therapy.
- Administration of the compositions can be via any of the accepted modes of administration for agents that serve similar utilities.
- While human dosage levels have yet to be optimized for the compounds described herein, generally, a daily dose is from about 0.01 to 15.0 mg/kg of body weight, preferably about 0.1 to 7.5 mg/kg of body weight, and most preferably about 0.3 to 1.5 mg/kg of body weight. Thus, for administration to a 70 kg person, the dosage range would be about 0.7 to 1,000 mg per day, preferably about 7.0 to 500 mg per day, and most preferably about 21 to 100 mg per day. The amount of active compound administered will, of course, be dependent on the subject and disease state being treated, the severity of the affliction, the manner and schedule of administration and the judgment of the prescribing physician.
- Compositions described herein may be employed In any skin care application where decreased Inflammatory response is desirable. For example, the compositions may be incorporated Into leave-on and rinse-off acne preparations, facial milks and conditioners, shower gels, foaming and non-foaming facial cleansers, cosmetics, hand and body lotions, leave-on moisturizers, cosmetic and cleaning wipes, salves for poison ivy, chicken pox, pruritus, or the like. Generally, for dermal applications, topical administration is preferred; however, systemic administration, as described elsewhere herein, is also possible.
- In employing the compositions for treatment of the above conditions, any therapeutically acceptable mode of administration can be used. The redox active molecules can be administered either alone or in combination with other therapeutically acceptable excipients, including solid, semi-solid, liquid or aerosol dosage forms, such as, for example, tablets, capsules, powders, liquids, suspensions, suppositories, aerosols or the like. The compositions can also be administered in sustained or controlled release dosage forms, including depot injections, osmotic pumps, pills, transdermal (including electro-transport) patches, and the like, for the prolonged administration of the compound at a predetermined rate, preferably In unit dosage forms suitable for single administration of precise dosages. The compositions will typically include a conventional pharmaceutical carrier or excipient, and a redox-active molecule. In addition, these compositions may include other medicinal agents, pharmaceutical agents, carriers, adjuvants, and the like, including, but not limited to anticoagulants, blood clot dissolvers, permeability enhancers and slow release formulations.
- Generally, depending on the intended mode of administration, the therapeutically acceptable composition will contain about 0.1% to 90%, preferably about 0.5% to 50%, by weight of a redox-active molecule, the remainder being suitable pharmaceutical excipients, carriers, etc.
- One preferred manner of administration for the conditions detailed above is oral, using a convenient daily dosage regimen which can be adjusted according to the degree of affliction. For such oral administration, a therapeutically acceptable, non-toxic composition is formed by the incorporation of any of the normally employed excipients, such as, for example, mannitol, lactose, starch, magnesium stearate, sodium saccharine, talcum, cellulose, sodium crosscarmellose, glucose, gelatin, sucrose, magnesium carbonate, and the like. Such compositions take the form of solutions, suspensions, tablets, dispersible tablets, pills, capsules, powders, sustained release formulations and the like.
- Preferably the compositions will take the form of a pill or tablet and thus the composition will contain, along with the active ingredient, a diluent such as lactose, sucrose, dicalcium phosphate, or the like; a lubricant such as magnesium stearate or the like; and a binder such as starch, gum acacia, polyvinylpyrrolidine, gelatin, cellulose and derivatives thereof, and the like.
- Liquid therapeutically administrable compositions can, for example, be prepared by dissolving, dispersing, etc. a redox-active compound and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline, aqueous dextrose, glycerol, glycols, ethanol, and the like, to thereby form a solution or suspension. If desired, the pharmaceutical composition to be administered may also contain minor amounts of nontoxic auxiliary substances such as wetting agents, emulsifying agents, or solubilizing agents, pH buffering agents and the like, for example, sodium acetate, sodium citrate, cyclodextrine derivatives, sorbitan monolaurate, triethanolamine acetate, triethanolamine oleate, etc. Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania, 15th Edition, 1975. The composition or formulation to be administered will, in any event, contain a quantity of the active compound in an amount effective to alleviate the symptoms of the subject being treated.
- Dosage forms or compositions containing active ingredient in the range of 0.005% to 95% with the balance made up from non-toxic carrier may be prepared.
- For oral administration, a therapeutically acceptable non-toxic composition is formed by the incorporation of any of the normally employed excipients, such as, for example pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, talcum, cellulose derivatives, sodium crosscarmellose, glucose, sucrose, magnesium carbonate, sodium saccharin, talcum and the like. Such compositions take the form of solutions, suspensions, tablets, capsules, powders, sustained release formulations and the like. Such compositions may contain 0.01%-95% active ingredient, preferably 0.1-50%.
- For a solid dosage form, the solution or suspension, in for example propylene carbonate, vegetable oils or triglycerides, is preferably encapsulated in a gelatin capsule. Such diester solutions, and the preparation and encapsulation thereof, are disclosed in
U.S. Patents Nos. 4,328,245 ;4,409,239 ; and4,410,545 . For a liquid dosage form, the solution, e.g. in a polyethylene glycol, may be diluted with a sufficient quantity of a therapeutically acceptable liquid carrier, e.g. water, to be easily measured for administration. - Alternatively, liquid or semi-solid oral formulations may be prepared by dissolving or dispersing the redox-active compound in vegetable oils, glycols, triglycerides, propylene glycol esters (e.g. propylene carbonate) and the like, and encapsulating these solutions or suspensions in hard or soft gelatin capsule shells.
- Other useful formulations include those set forth in
U.S. Patents Nos. Re. 28,819 and4,358,603 . - The formulation can be administered in a single unit dosage form for continuous treatment or in a single unit dosage form ad libitum when relief of symptoms is specifically required. For example, the formulation may be administered as a bolus or as a continuous intravenous infusion after onset of symptoms of stroke, myocardial infarction or chronic heart failure.
- Parenteral administration is generally characterized by injection, either subcutaneously, intramuscularly or intravenously. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol or the like. In addition, if desired, the pharmaceutical compositions to be administered may also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, solubility enhancers, and the like, such as for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate, cyclodextrins, etc.
- Another devised approach for parenteral administration employs the implantation of a slow-release or sustained-release system, such that a constant level of dosage is maintained. See, e.g.,
U.S. Patent No. 3,710,795 . The percentage of active compound contained in such parenteral compositions is highly dependent on the specific nature thereof, as well as the activity of the compound and the needs of the subject. However, percentages of active ingredient of 0.01% to 10% in solution are employable, and will be higher if the composition is a solid which will be subsequently diluted to the above percentages. Preferably the composition will comprise 0.2-2% of the active agent in solution. Nasal solutions of the active compound alone or in combination with other therapeutically acceptable excipients can also be administered. - The cyclic voltammetry measurement was conducted in a three-electrode system comprising a microelectrode made from platinum (a disk of 1.6mm2 area), a counter electrode made from a coiled platinum wire and a silver/silver chloride reference electrode, on a voltammetric analyzer Epsilon with a C-3 cell , all from Bioanalytical Systems (West Lafayette, IN).
- A solution of tetrabutyl ammonium perchlorate (TBAP) (500 mM final concentration) in N, N-dimethyl formamide (DMF) (5 mL) was prepared in a graduated flask. The compound was added to reach a concentration of 10 mM. The solution was stirred and sonicated if necessary to make sure that the compound was fully dissolved.
- The reference electrode was kept in a 3 M solution of NaCl in distilled water. The reference electrode was rinsed with water, then methanol, and dried by air blow. The reference electrode was gently shaken, if necessary, to remove any air bubble within the base.
- The auxiliary electrode was rinsed with water, then methanol and dried by air blow.
- The working electrode was polished on a nylon pad coated with a 1 µ-meter diamond slurry in oil, then rinsed thoroughly with methanol. It is then polished on a corduroy pad coated with a 0.15 µ-meter alumina slurry in water, then rinsed thoroughly with water and methanol, then dried by air blow.
- Prior to beginning a measurement and when changing compound solutions, the electrochemical cell was thoroughly cleaned with methanol, including glass reservoir, Teflon cap, and purge lines. Likewise, all electrodes were cleaned with methanol upon removal for any reason, including prior to polishing the working electrode between runs. While the working electrode was being polished, the reference electrode was removed from the test solution, rinsed with methanol and water, and stored in aqueous 3M NaCl solution.
-
Reduction Oxidation Initial potential: 0 mV Initial potential: 0 mV Switching potential #1: -1800 mV Switching potential #1: 1800 mV Switching potential #2: 1800 mV Switching potential #2: -1800 mV Final potential: -800 Final potential: 800 Number of segments: 3 Number of segments: 3 Scan rate: 100 mV/s, 2000mV/s, 20 mV/s Scan rate: 100 mV/s, 2000mV/s, 20 mV/s - Other parameters could be investigated after the initial oxidative and reductive runs. The parameters of such runs are highly compound dependant and it is up to the judgment of those collecting data, but minimally the above parameters should be used.
- The solution containing the compound was transferred into the cell, where the three electrodes were immersed. The solution was stirred and purged with dry argon for at least 1 minute to remove any oxygen, after which a blanket of argon was kept above the solution to prevent any diffusion of oxygen. Stirring was halted prior to collecting data. Care was taken to ensure there were no bubbles attached to any of the electrodes prior to beginning electrochemical measurements.
- The solution is not a perfect conductor, and its resistance R introduces a voltage drop when a current flows between the working to the auxiliary electrode. This voltage drop introduces an error between the nominal voltage imposed by the voltammetric analyzer and the actual voltage between the working and auxiliary electrodes. The instrument can automatically measure and compensate for this iR drop, using the "iR Compensation" feature of the software. Normal values are the following:
- Cell resistance R < 1000 ohms
- RC constant >10 µs
- % resistance to be compensated: 50 to 100%
- Uncompensated resistance < (R - 300 ohms)
- The cyclic voltammogram was recorded. After each cyclic voltammogram was recorded, the working electrode was polished again, as indicated above, to regenerate a clean, electroactive surface.
- The square wave voltammetry measurement was conducted in a three-electrode system comprising a microelectrode made from platinum (a disk of 1.6mm2 area), a counter electrode made from coiled platinum wire and a silver/silver chloride reference electrode, on a voltammetric analyzer CV-50W with a C-3 cell , all from Bioanalytical Systems (West Lafayette, IN).
- A solution of tetrabutyl ammonium perchlorate (TBAP) (500 mM final concentration) in N,N-dimethyl formamide (DMF) (5 mL) is prepared in a graduated flask. The compound is added to reach an arbitrary concentration, 10 mM is desired but less is acceptable. This method is especially useful for low concentration analytes, those molecules that are difficult to dissolve in DMF or available only in small quantities. The solution is stirred and sonicated if necessary to make sure that the compound is dissolved to the best of its ability.
- The reference electrode is kept in a 3 M solution of NaCl in distilled water. The reference electrode is rinsed with water, then methanol, and dried by air blow. The reference electrode is gently shaken, if necessary, to remove any air bubble at the base. The auxiliary electrode is rinsed with water, then methanol and dried by air blow.
- The working electrode is polished on a nylon pad coated with a 1 µ-meter diamond slurry in oil, then rinsed thoroughly with methanol. It is then polished on a corduroy pad coated with a 0.15 µ-meter alumina slurry in water, then rinsed thoroughly with water and methanol, then dried by air blow.
- Prior to beginning and when changing compound solutions, the electrochemical cell should be thoroughly cleaned with methanol including the glass reservoir, Teflon cap, and purge lines. Likewise, all electrodes should be promptly cleaned with methanol upon removal for any reason, including prior to polishing the working electrode between runs. While the working electrode is being polished, the reference electrode should be removed from the test solution, rinsed with methanol and water, and stored in aqueous 3M NaCl solution.
-
Reduction Oxidation Initial potential: 1800 mV Initial potential: -1800 mV Switching potential #1: -1800 mV Switching potential #1: 1800 mV Step Potential: 4 Step Potential: 4 Amplitude: 25 Amplitude: 25 Frequency: 15 Frequency: 15 - Other parameters could be investigated after the initial oxidative and reductive runs. The parameters of such runs are highly compound dependent and it is up to the judgment of those collecting data, but minimally the above parameters should be used.
- The solution containing the compound is transferred into the cell, where the three electrodes are immersed. The solution is stirred and purged with dry argon for at least 1 minute to remove any oxygen, after which a blanket of argon is kept above the solution to prevent any diffusion of oxygen. Stirring is halted prior to collecting data. Care must be taken to ensure there are no bubbles attached to any of the electrodes prior to beginning electrochemical measurements.
- The voltammogram can then be recorded. After each square wave voltammogram is recorded, the working electrode is polished again, as indicated above, to regenerate a clean, electroactive surface.
- Endothelial-Leukocyte Adhesion Molecule (ELAM), also known as E-selectin, is expressed on the surface of endothelial cells. In this assay, lipopolysaccharide (LPS) and IL-1β were used to stimulate the expression of ELAM; test agents were tested for their abilities to reduce this expression, in accordance with studies showing that reduction of leukocyte adhesion to endothelial cell surface was associated with decreased cellular damage (e.g., Takada, M. et al., Transplantation 64: 1520-25, 1997; Steinberg, J.B. et al., J. Heart Lung Trans. 13:306-313, 1994).
- Endothelial cells may be selected from any of a number of sources and cultured according to methods known in the art; including, for example, coronary artery endothelial cells, human brain microvascular endothelial cells (HBMEC; Hess, D.C. et al., Neurosci. Lett. 213(1): 37-40, 1996), or lung endothelial cells. Cells were conveniently cultured in 96-well plates. Cells were stimulated by adding a solution to each well containing 10 µg/ml LPS and 100 µg/ml IL-1β for 6 hours in the presence of test agent (specific concentrations and time may be adjusted depending on the cell type). Treatment buffer was removed and replaced with pre-warmed Fixing Solution® (100 µL/well) for 25 minutes at room temperature. Cells were then washed 3X, then incubated with Blocking Buffer (PBS + 2% FBS) for 25 minutes at room temperature. Blocking Buffer containing Monoclonal E-Selectin Antibody (1:750, Sigma Catalog #S-9555) was added to each well. Plates were sealed and stored at 4° overnight. Plates were washed 4X with 160 µL Blocking Buffer per well. Second Antibody-HRP diluted 1:5000 in Blocking Buffer was then added (100 µL/well), and plates were incubated at room temperature (protected from light) for two hours. Plates were then washed 4X with Blocking Buffer before addition of 100 µL of ABTS Substrate solution at room temperature (Zymed, Catalog #00-2024). Wells were allowed to develop for 35 minutes, before measurement at 402 nm in a Fluoroskan® Reader with shake program for 10 seconds. Positive results were recorded as a decrease in ELAM concentration in tested wells, as compared to control wells.
- This assay was used to induce ischemia by anoxia-reoxygenation in cultured hippocampal neuronal cells. Test compounds were added to assess potency and efficacy against ischemia-induced neuronal cell injury and cell death.
- Neurobasal/B27i: Neurobasal medium (available from Invitrogen, San Diego, CA) with 1x B27 supplement (Invitrogen), 0.5 µM L-glutamine, 25 µM L-glutamic acid, and 1 x Penicillin/Streptomycin.
- Hank's Basic Salt Solution (HBSS, Ca/Mg-free) is prepared by preparing 1X Hanks CMF (Gibco) supplemented with HEPES (10 mM, pH 7.3), sodium bicarbonate (0.35%), 1X Penicillin/Streptomycin, and 1 mM MEM sodium pyruvate.
- Poly-D-lysine (Sigma, St. Louis, MO), 50 µg/ml solution.
- Sigmacote (Sigma, St. Louis, MO).
- Plastic Culture Flasks (T75 cm2) or 24-well cell culture plates treated with Poly-D-Lysine (Sigma, St. Louis, MO).
- A pregnant female mouse (E18-E19) was euthanized with CO2 followed by removal of the uterus, which was then placed in a sterile plastic petri dish. The embryos were removed from the sac, and the embryonic brains removed and immersed in cold (4°C) Buffered Salt Solution (HBSS; Ca/Mg free; Life Technologies) in a small petri dish. Hippocampi were then removed from the brains under a dissecting microscope and placed on a paraffin-covered dish. The meninges were stripped away and the dissected hippocampi were collected in a small petri dish in HBSS. The hippocampi were transferred to a 15-ml centrifuge tube (normally 10-12 brains) filled with HBSS. The tube containing the brains was centrifuged at 1000 rpm for 2 min in a tabletop centrifuge. The supernatant was removed, 2 ml of HBSS was added to the hippocampi in the tube, and the resulting suspension was triturated 2 times each with long-tipped siliconized glass pipettes having progressively smaller apertures, starting with a pipette with a standard size opening (approximately 1.0 mm diameter), following with one having an aperture of half standard size (approximately 0.5 mm diameter), then with one having an aperture about one-half that size (0.25 mm diameter). The suspension was then centrifuged again at 1000 rpm for 2 min in a tabletop centrifuge, the supernatant was discarded, and 2 ml of Neurobasal/B27i (with antibiotics) was added to the tube. The trituration procedure described above was then repeated on this suspension.
- The density of cells was determined on a small aliquot of cells using standard counting procedures and correcting for cell viability by trypan blue stain exclusion. Using this procedure, the expected yield is 3 × 105- 6 × 105 cells/brain. Cells were then added to PDL-coated 24 well plates, flasks or MetTek dishes in Neurobasal/B271 at a density of about 1.5 × 106 cells (T75 flask) or about 100,000 cells/well of a 24-well plate. Plated cells were incubated at 37°C in an atmosphere of 5 % CO2/ 95 % O2. Media was renewed after 3-4 days by replacing half of it with fresh Neurobasal/B27m medium, containing 5 µM cytosine arabinoside (Ara-C). Seven to eight days from the initial culture, the media was renewed again, by removing one-half or it and replacing with an equal amount of fresh Neurobasal/B27m medium (without Ara-C).
- Neurobasal media, NoG neurobasal media, B27 supplement and B27 Supplement minus AO (Invitrogen).
- Neurobasal/B27 medium is prepared with 2X B27 minus AO supplement, 0.5 mM L-glutamine and 0.25X penicillin/streptomycin.
- Cell Tracker Green was obtained from Molecular Probes and a fresh 5µM solution was prepared from 10 mM stock just before use.
- NoG-Neurobasal contains NoG neurobasal medium plus 0.5 mM glucose, 0.1 mM L-glutamine and 0.25X Penicillin/Streptomycin.
- Primary hippocampal neuronal cells were prepared according to the methods described above and were cultured in poly-D-lysine coated 24 well plates for 10-11 days prior to use.
- Deoxygenated LoG-Neurobasal medium (100 ml) was prepared by pre-equilibrating the medium in a T150 cm2 flask in a hypoxic chamber overnight. Following pre-incubation under hypoxic conditions, the LoG-Neurobasal media was lightly bubbled with 100% N2 for 30 min to completely deoxygenate the media. An additional 20 ml LoG-Neurobasal was pre-equilibrated in a T75 cm2 flask and 100 ml Neurobasal/B27AO was incubated in a normal incubator (5% CO2 overnight. Reoxygenated medium was prepared by placing medium overnight in the culture incubator (5% CO2/95% O2) prior to use.
- Existing culture medium (Neurobasal/B27m) was removed from the cells by aspiration. Cells were washed once with 2 ml/well (24-well culture plates) of glucose free-BSS. Neurons were replenished 10-11 days after initial culture with deoxygenated LoG-Neurobasal (1 ml per well for each well of a 24-well plate). Test compounds were added directly to each well (3 concentrations of the compound plus positive control, each in triplicate). Most test compounds were dissolved in 100% DMSO; concentrations were adjusted such that the final concentration of DMSO in the cell media never exceeded 0.5%. Plates containing cells with test compounds were placed in a hypoxic chamber for 5hr with plate lids ajar. For normoxia controls, pre-equilibrated normoxic LoG-Neurobasal medium was added to each well of cells, and the plate was replaced in the normal culture incubator for 5 hr. After 5 hr of hypoxia, the existing media was carefully aspirated off, and 2mL of new, re-oxygenated (pre-equilibrated) Neurobasal/B27AO was added to each well. The same test compounds (in the same the concentrations) were added back into the corresponding wells. Plates were placed in the cell culture incubator (5% CO2/95% O2) and reoxygenated for 20-24 hr. After reoxygenation for 20-24 hr, live neurons were quantitated using the cell tracker green fluorescence method, described below.
- To test for cell viability, existing culture medium was aspirated from each well of the 24 well plates, and neurons were washed once with 2 ml of HBSS (pH 7.4, pre-warmed to 30-37°C). To each well was added one milliliter of 5 µM Cell Tracker Green fluorescent dye dissolved in HBSS. Plates were placed in the dark at room temperature for 15 minutes, and are then washed with two milliliters of HBSS. One milliliter of HBSS was then added to each well, and fluorescent cells were counted using a fluorescent microscope. Significantly increased cell viability compared to control cells was indicative of a protective compound.
- This procedure was used to induce high glutamate-induced oxidative stress (HGOS) in a dopaminergic neuronal cell line. Using this assay the potency and efficacy of test articles against HGOS neuronal cell injury and cell death can be established in a high throughput manner.
-
- Dopaminergic neuronal cell lines
- DMEM-No Glucose (Life Technologies Cat # 11966-025)
- L-glutamine (Life Technologies Cat # 25030-081)
- L-glutamic acid, monosodium salt (Sigma Cat # G5889)
- D-glucose (Sigma Cat # G-6151)
- 10x HBSS buffer(pH 7.4) (950ml Pyrogen-free water, 2.44g/L MgCl2.6H20, 3.73g/L KCl, 59.58g/L Hepes, 58.44g/L NaCl, 1.36g/L KH2PO4, 1.91 g/L CaCl2.2H2O and pH to 4.5 with HCl)
- Cell Tracker Green fluorescent dye (Molecular Probes, Cat # 2925). Prepare a 5µM solution in pre-warmed HBSS just prior to use.
- Sterile 96-well plates precoated with poly-D-lysine (Corning Catalog # 3665)
- 96-well deep well mother plate, DyNA Block 1000 (VWR Catalog # 40002-008)
- The cells were seeded into 96-well plates at a density of 2000 per well and left to grow for 72 hours in a 33°C incubator with 5% CO2 in air atmosphere. The passage number of the cells for each assay experiment were no later than p11 in order to minimize experimental variation.
-
VWRBrand DyNA Block 1000, deep well mother plates (VWR Cat. # 40002-008) were used for the preparation of the test compounds. - All compounds were dissolved in DMEM-No Glu containing 1mM glucose, 30 mM glutamate and 1x Pen/Strep. DMEM-No Glu with 1mM glucose and 1x P/S was used as the negative control, DMEM-No Glucose with 1 mM glucose, 100 M glutamate was used as a positive control and 100µM Glutathione was added to the positive control as a standard. All of the procedures for this involving the making and dilution of compounds were performed using aseptic conditions and with minimal light.
- The plates were removed from the incubator and examined under the microscope for morphological appearance and density. Using an aseptic technique and an 8-channel aspirator the media was carefully removed from the cells and replaced with 200µl of 1x HBSS. This was done as quickly as possible to prevent the cells drying out. The plates were then placed in the humidified 37°C incubators of the
Biomek 2000 Side Loader. Four plates were washed at a time so as to minimize the time that the cells were sitting in 1x HBSS prior to addition of the compound test solution. - The Beckman Biomek workstations were used to load the compounds and controls from the mother plates onto the cell plates that were prewashed with HBSS under sterile conditions. The plates were incubated in the upper HTS incubator at 37°C in 5% CO2 for exactly 16 hrs. The following day, using the Beckman Biomek workstations, the plates were removed from the incubator. Using Cell Tracker Addition, the compounds were removed from the plates, washed once with 200µM of pre-warmed 1x HBSS and then 100µL of 5µM Cell Tracker Green was added to each well. The plates were incubated at 37°C for 30 min to allow the dye to enter the cell and be cleaved by the esterases. After washing the cells twice with prewarmed 1x HBSS, the plates were read with the 485 excitation; 538 emission filter pair on a Fluoroskan.
Claims (18)
- A method of identifying and selecting therapeutic compounds having a predetermined core structure, said method comprising:establishing a relationship between physical-chemical profile and biological activity; wherein the physical-chemical profile comprises one or more parameters selected from onset of oxidation, potential of oxidation, potential of reduction, reversibility of oxidation, reversibility of reduction, current of oxidation or current of reduction; and wherein the biological activity is measured in an assay effective in detecting compounds for the treatment of a targeted disorder;testing further potential therapeutic candidates with said core structure for their physical- chemical properties; andselecting therapeutic compounds based on their physical-chemical parameters falling within a range predefined by the physical-chemical/biological relationship of the previously tested subset of compounds,wherein the predetermined core structure is selected from a quinone core structure, the targeted disorder is oxidative stress, inflammation, ischemia, neurodegeneration, or Alzheimer's disease, and the physical-chemical profile of the compounds comprises a parameter for the total reversibility of reduction of 75% or more, said parameter for reversibility of reduction being a ratio of reduction wave maximum current to oxidation wave maximum current of 0.75 or more;or wherein the predetermined core structure is selected from a core structure of Formula I:
(I-a) additional substitution at the phenyl rings of the core structure of Formula I does not include a nitro group; and the physical-chemical profile of the compounds comprises the parameter for potential of the first oxidation wave where said potential falls below 1000mV versus a silver/silver chloride reference electrode; or
(I-b) additional substitution at the phenyl rings of the core structure of Formula I includes a nitro group and the physical-chemical profile of the compounds comprises one or more parameters selected from the parameter for potential of the first oxidation wave where said potential falls between 950mV and 1250mV versus a silver/silver chloride reference electrode and the parameter for reversibility of the first oxidation wave measuring more than 20%, said parameter for reversibility of the first oxidation wave being a ratio of first oxidation wave maximum current to reduction wave maximum current of 0.2 or more;
or wherein the predetermined core structure is selected from a core structure of Formula III:
(III-a) the physical-chemical profile of the compounds comprises the parameter for potential of the first oxidation wave where said potential falls under 1250mV versus a silver/silver chloride reference electrode; or
(III-b) the physical-chemical profile of the compounds comprises the parameter for potential of the first reduction where said potential is more negative than -790mV versus a silver/silver chloride reference electrode
or wherein the predetermined core structure is selected from a core structure of Formula IV: - The method of claim 1, wherein said physical-chemical profile comprises a cyclic voltammetric profile.
- The method of claim 1, wherein the physical-chemical profile comprises the parameter for onset of oxidation.
- The method of claim 1, wherein the physical-chemical profile comprises the parameter for potential of oxidation wave.
- The method of claim 1, wherein the physical-chemical profile comprises the parameter for reversibility of one or more oxidation waves.
- The method of claim 1, wherein the physical-chemical profile comprises the parameter for reversibility of one or more reduction waves.
- The method of claim 1, wherein the physical-chemical profile comprises the parameter for potential of reduction wave.
- The method of any one of the preceding claims, wherein the assay is a cell-based assay comprising one or more assays selected from the High Glutamate-induced Oxidative Stress (HGOS) assay wherein the compounds in the previously tested subset group of compounds have the ability to protect at least 30% of energetically competent cells against stressor induced cell death; and the E-selectin (ELAM) assay wherein the compounds in the previously tested subset group of compounds exhibit an EC50 lower than 30 µM.
- The method of claim 8, wherein the therapeutic compound is selected if it comprises a stilbene core structure and if its physical-chemical profile comprises one or more parameters selected from the parameter for potential of the first oxidation wave where said potential falls between 800mV and 1400mV versus a silver/silver chloride reference electrode, and the parameter for the reversibility of the first oxidation wave that measures 20% or more.
- The method of claim 8, wherein the therapeutic compound is selected if it comprises a core structure of Formula I:(a) additional substitution at the phenyl rings does not include a nitro group; and if its physical-chemical profile comprises the parameter for potential of the first oxidation wave where said potential falls below 1000mV versus a silver/silver chloride reference electrode; or(b) additional substitution at the phenyl rings includes a nitro group; and if its physical-chemical profile comprises one or more parameters selected from the parameter for potential of the first oxidation wave where said potential falls between 950mV and 1250mV versus a silver/silver chloride reference electrode and the parameter for reversibility of the first oxidation wave measuring more than 20%.
- The method of claim 3, wherein the physical-chemical profile comprises the parameter for onset of oxidation and the assay comprises the E-selectin (ELAM) cell based assay detecting compounds with an EC50 lower than 30µM.
- The method of claim 4, wherein the physical-chemical profile comprises the parameter for potential of oxidation wave and the cell based assay comprises the hippocampal anoxia reoxygenation cell death assay protecting at least 30% of the cells against stressor induced cell death.
- The method of claim 4, wherein the physical-chemical profile comprises the parameter for potential of the first oxidation wave and the assay is the Thioflavin T binding assay measuring reduction of amyloid-β fibril formation.
- The method of claim 13, wherein the compound is selected if it comprises an apomorphine core of Formula III:(a) the physical-chemical profile comprises the parameter for potential of the first oxidation wave where said potential falls under 1250mV versus a silver/silver chloride reference electrode; or(b) the physical-chemical profile comprises the parameter for potential of the first reduction where said potential is more negative than -790mV versus a silver/silver chloride reference electrode.
- The method of claim 6, wherein the physical-chemical profile comprises the parameter for reversibility of reduction wave and the assay comprises the E- selectin (ELAM) cell based assay detecting compounds with an EC50 lower than 30µM.
- The method of claim 15, wherein the therapeutic compound is selected if it comprises a quinone core structure and if its physical-chemical profile comprises a parameter for the total reversibility of reduction of 75% or more.
- The method of claim 12, wherein the therapeutic compound is selected if it comprises a chroman core structure of Formula IV,
- The method of any one of the preceding claims, wherein the physical-chemical profile additionally comprises one or more parameters selected from energy profile parameters and transport profile parameters.
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US42272702P | 2002-10-30 | 2002-10-30 | |
US422727P | 2002-10-30 | ||
US48773403P | 2003-07-16 | 2003-07-16 | |
US487734P | 2003-07-16 | ||
PCT/US2003/034420 WO2004042353A2 (en) | 2002-10-30 | 2003-10-29 | Identifying therapeutic compounds based on their physical-chemical properties |
Publications (3)
Publication Number | Publication Date |
---|---|
EP1558922A2 EP1558922A2 (en) | 2005-08-03 |
EP1558922A4 EP1558922A4 (en) | 2008-08-06 |
EP1558922B1 true EP1558922B1 (en) | 2013-02-27 |
Family
ID=32314462
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP03781483A Expired - Lifetime EP1558922B1 (en) | 2002-10-30 | 2003-10-29 | Identifying therapeutic compounds based on their physical-chemical properties |
Country Status (6)
Country | Link |
---|---|
US (3) | US7491312B2 (en) |
EP (1) | EP1558922B1 (en) |
JP (1) | JP4599292B2 (en) |
AU (2) | AU2003287250B9 (en) |
CA (1) | CA2504334A1 (en) |
WO (1) | WO2004042353A2 (en) |
Families Citing this family (33)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1558922B1 (en) * | 2002-10-30 | 2013-02-27 | Ampere Life Sciences, Inc. | Identifying therapeutic compounds based on their physical-chemical properties |
WO2005032544A1 (en) | 2003-09-19 | 2005-04-14 | Galileo Pharmaceuticals, Inc. | Treatment of mitochondrial diseases |
US8088773B2 (en) | 2005-05-12 | 2012-01-03 | The Texas A&M University System | Therapeutic compositions and methods |
EP2471530B1 (en) | 2005-06-01 | 2017-01-11 | Edison Pharmaceuticals, Inc. | Redox-active therapeutics for treatment of mitochondrial diseases and other conditions and modulation of energy biomarkers |
KR101431279B1 (en) | 2005-07-29 | 2014-08-20 | 리스버로직스 코퍼레이션 | Pharmaceutical compositions for the prevention and treatment of complex diseases and their delivery by insertable medical devices |
CA2635280C (en) | 2006-02-22 | 2017-12-12 | Edison Pharmaceuticals, Inc. | Side chain variants of redox-active therapeutics for treatment of mitochondrial diseases and other conditions and modulation of energy biomarkers |
WO2009061744A2 (en) | 2007-11-06 | 2009-05-14 | Edison Pharmaceuticals, Inc. | 4- (p-quinonyl)-2-hydroxybutanamide derivatives for treatment of mitochondrial diseases |
EA023618B1 (en) | 2008-01-08 | 2016-06-30 | Эдисон Фармасьютикалз, Инк. | (HET)ARYL-p-QUINONE DERIVATIVES FOR TREATMENT OF MITOCHONDRIAL DISEASES |
EP2262508B1 (en) | 2008-03-05 | 2018-10-03 | BioElectron Technology Corporation | SUBSTITUTED-p-QUINONE DERIVATIVES FOR TREATMENT OF OXIDATIVE STRESS DISEASES |
US8716486B2 (en) * | 2008-06-25 | 2014-05-06 | Edison Pharmaceuticals, Inc. | 2-heterocyclylaminoalkyl-(p-quinone) derivatives for treatment of oxidative stress diseases |
WO2010030607A1 (en) | 2008-09-10 | 2010-03-18 | Edison Pharmaceuticals, Inc. | Treatment of pervasive developmental disorders with redox-active therapeutics |
US10039722B2 (en) | 2008-10-14 | 2018-08-07 | Bioelectron Technology Corporation | Treatment of oxidative stress disorders including contrast nephropathy, radiation damage and disruptions in the function of red cells |
SI2963006T1 (en) | 2008-10-28 | 2019-01-31 | Bioelectron Technology Corporation | Composition containing alpha-tocotrienol quinone,and intermediates thereof |
DK2424495T3 (en) | 2009-04-28 | 2018-04-30 | Bioelectron Tech Corp | TREATMENT OF LEVERS HEREDIC OPTICAL NEUROPATHY AND DOMINANT OPTICAL ATROPHY WITH TOCOTRIENOL QUINONES |
CN102647981B (en) * | 2009-08-26 | 2016-09-28 | 爱迪生制药有限公司 | Prevention and the method for the treatment of cerebral ischemia |
JP2011106954A (en) * | 2009-11-17 | 2011-06-02 | Mitsui Eng & Shipbuild Co Ltd | Monitoring method of concentration of polyhydric phenol |
WO2011113018A1 (en) * | 2010-03-12 | 2011-09-15 | Ampere Life Sciences, Inc. | Measurement and control of biological time |
EP2720689A4 (en) | 2011-06-14 | 2014-11-26 | Edison Pharmaceuticals Inc | Catechol derivatives for treatment of oxidative stress diseases |
JP2014520894A (en) | 2011-07-19 | 2014-08-25 | エジソン ファーマシューティカルズ, インコーポレイテッド | A method for the selective oxidation of alpha tocotrienol in the presence of non-alpha tocotrienol |
US9254287B2 (en) | 2012-06-11 | 2016-02-09 | Macuclear, Inc. | Therapeutic formulation and methods of treatment |
WO2014039862A1 (en) * | 2012-09-07 | 2014-03-13 | Edison Pharmaceuticals, Inc. | Quinone derivatives for use in the modulation of redox status of individuals |
JP2016515526A (en) | 2013-03-15 | 2016-05-30 | エジソン ファーマシューティカルズ, インコーポレイテッド | Alkyl-heteroaryl substituted quinone derivatives for the treatment of oxidative stress disorders |
US9868711B2 (en) | 2013-03-15 | 2018-01-16 | Bioelectron Technology Corporation | Phenazine-3-one and phenothiazine-3-one derivatives for treatment of oxidative stress disorders |
US9296712B2 (en) | 2013-03-15 | 2016-03-29 | Edison Pharmaceuticals, Inc. | Resorufin derivatives for treatment of oxidative stress disorders |
US9670170B2 (en) | 2013-03-15 | 2017-06-06 | Bioelectron Technology Corporation | Resorufin derivatives for treatment of oxidative stress disorders |
KR102626895B1 (en) | 2014-12-16 | 2024-01-18 | 피티씨 테라퓨틱스, 인크. | Polymorphic and amorphous forms of (r)-2-hydroxy-2-methyl-4-(2,4,5-trimethyl-3,6-dioxocyclohexa-1,4-dienyl)butanamide |
US10702571B2 (en) | 2015-12-03 | 2020-07-07 | The University Of North Carolina At Pembroke | Materials for cathepsin B enhancement and methods of use |
WO2017106786A1 (en) | 2015-12-16 | 2017-06-22 | Bioelectron Technology Corporation | Improved methods for enriching alpha-tocotrienol from mixed tocol compositions |
WO2017106803A1 (en) | 2015-12-17 | 2017-06-22 | Bioelectron Technology Corporation | Flouroalkyl, flouroalkoxy, phenoxy, heteroaryloxy, alkoxy, and amine 1,4-benzoquinone derivatives for treatment of oxidative stress disorders |
JP2018083799A (en) | 2016-11-15 | 2018-05-31 | バイオエレクトロン テクノロジー コーポレイション | 2-SUBSTITUTED AMINO-NAPHTHO[1,2-d]IMIDAZOLE-5-ONE COMPOUND OR PHARMACEUTICALLY ALLOWABLE SALTS THEREOF |
US11467124B2 (en) * | 2017-05-02 | 2022-10-11 | The University Of British Columbia | Apparatus, systems and methods for in situ measurement of an oxidation / reduction potential and pH of a solution |
CN113365616A (en) | 2018-10-17 | 2021-09-07 | Ptc医疗公司 | 2,3,5-trimethyl-6-nonylcyclohexa-2,5-diene-1, 4-diones for the inhibition and treatment of alpha-synucleinopathies, tauopathies and other diseases |
AU2022306868A1 (en) | 2021-07-08 | 2024-02-22 | Ptc Therapeutics, Inc. | Pharmaceutical compositions comprising 2,3,5-trimethyl-6-nonylcyclohexa-2,5-diene-1,4-dione |
Family Cites Families (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4997526A (en) | 1985-03-19 | 1991-03-05 | Eic Laboratories, Inc. | Assaying for a biologically active component |
JP3251582B2 (en) | 1991-02-04 | 2002-01-28 | ケイエス・バイオメディックス・リミテッド | Naphthoquinone derivatives for treating chronic inflammation |
DE69433935D1 (en) | 1993-11-26 | 2004-09-09 | Lawrence B Hendry | THE DESIGN OF DRUGS THAT INTERRUPT THE RECEPTOR LIGAND DNA INTERACTIONS |
US5463564A (en) | 1994-09-16 | 1995-10-31 | 3-Dimensional Pharmaceuticals, Inc. | System and method of automatically generating chemical compounds with desired properties |
SI0801564T1 (en) | 1994-12-28 | 2002-08-31 | Janssen Pharmaceutica N.V. | Use of nebivolol as an anti-atherogenic |
US5716928A (en) | 1995-06-07 | 1998-02-10 | Avmax, Inc. | Use of essential oils to increase bioavailability of oral pharmaceutical compounds |
US5807708A (en) | 1996-07-30 | 1998-09-15 | Millennium Pharmaceuticals, Inc. | Conservin nucleic acid molecules and compositions |
IL129728A0 (en) | 1996-11-04 | 2000-02-29 | Dimensional Pharm Inc | System method and computer program product for the visualization and interactive processing and analysis of chemical data |
US6372772B1 (en) | 1997-08-01 | 2002-04-16 | Prolx Pharmaceuticals Corporation | Inhibitors of redox signaling and methods of using same |
US6344330B1 (en) | 1998-03-27 | 2002-02-05 | The Regents Of The University Of California | Pharmacophore recombination for the identification of small molecule drug lead compounds |
GB2360007A (en) * | 2000-03-10 | 2001-09-12 | Chen Yang Shiau | Welded heat sink/heat pipe for CPU |
US6387945B2 (en) | 2000-04-11 | 2002-05-14 | The Regents Of The University Of California | Lipoic acid analogs |
US20020034537A1 (en) | 2000-05-03 | 2002-03-21 | Brita Schulze | Cationic diagnostic, imaging and therapeutic agents associated with activated vascular sites |
US7182853B2 (en) | 2000-09-22 | 2007-02-27 | University Of Dayton | Redox control/monitoring platform for high throughput screening/drug discovery applications |
US20020132845A1 (en) * | 2000-12-15 | 2002-09-19 | Miller Guy Michael | Compositions and methods for the prevention and treatment of tissue ischemia |
WO2002053152A1 (en) | 2001-01-08 | 2002-07-11 | Imperial College Of Science, Technology & Medicine | Composition containing flavonoids for treatment brain disorders |
EP1558922B1 (en) * | 2002-10-30 | 2013-02-27 | Ampere Life Sciences, Inc. | Identifying therapeutic compounds based on their physical-chemical properties |
-
2003
- 2003-10-29 EP EP03781483A patent/EP1558922B1/en not_active Expired - Lifetime
- 2003-10-29 CA CA002504334A patent/CA2504334A1/en not_active Abandoned
- 2003-10-29 JP JP2005502210A patent/JP4599292B2/en not_active Expired - Fee Related
- 2003-10-29 WO PCT/US2003/034420 patent/WO2004042353A2/en active Search and Examination
- 2003-10-29 US US10/696,752 patent/US7491312B2/en not_active Expired - Fee Related
- 2003-10-29 AU AU2003287250A patent/AU2003287250B9/en not_active Ceased
-
2008
- 2008-12-16 US US12/336,449 patent/US20090162890A1/en not_active Abandoned
- 2008-12-24 US US12/344,025 patent/US20090163529A1/en not_active Abandoned
-
2009
- 2009-10-22 AU AU2009227912A patent/AU2009227912A1/en not_active Abandoned
Non-Patent Citations (2)
Title |
---|
KILMARTIN: "Electrochemical detection of natural antioxidants; principles and protocols", ANTIOXIDANTS AND REDOX SIGNALING, vol. 3, 2001, pages 941 - 955 * |
LASHUEL ET AL.: "New class of inhibitors of amyloid-beta fibril formation. Inplications for the mechanism of pathogenesis in Alzheimer diseases.", THE JOURNAL OF THE BIOLOGICAL CHEMISTRY, vol. 277, 2002, pages 42881 - 42890 * |
Also Published As
Publication number | Publication date |
---|---|
WO2004042353A3 (en) | 2004-08-19 |
US20040105817A1 (en) | 2004-06-03 |
WO2004042353A2 (en) | 2004-05-21 |
AU2009227912A1 (en) | 2009-11-12 |
AU2003287250A1 (en) | 2004-06-07 |
AU2003287250B2 (en) | 2009-07-23 |
US20090163529A1 (en) | 2009-06-25 |
JP2006517398A (en) | 2006-07-27 |
AU2003287250B9 (en) | 2010-01-28 |
JP4599292B2 (en) | 2010-12-15 |
US20090162890A1 (en) | 2009-06-25 |
EP1558922A4 (en) | 2008-08-06 |
CA2504334A1 (en) | 2004-05-21 |
US7491312B2 (en) | 2009-02-17 |
EP1558922A2 (en) | 2005-08-03 |
WO2004042353A8 (en) | 2005-07-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP1558922B1 (en) | Identifying therapeutic compounds based on their physical-chemical properties | |
Nadanaciva et al. | Mitochondrial impairment by PPAR agonists and statins identified via immunocaptured OXPHOS complex activities and respiration | |
Gobert et al. | Simultaneous quantification of serotonin, dopamine and noradrenaline levels in single frontal cortex dialysates of freely-moving rats reveals a complex pattern of reciprocal auto-and heteroreceptor-mediated control of release | |
Zou et al. | 2-NBDG as a fluorescent indicator for direct glucose uptake measurement | |
Liu et al. | Online electrochemical monitoring of dynamic change of hippocampal ascorbate: toward a platform for in vivo evaluation of antioxidant neuroprotective efficiency against cerebral ischemia injury | |
Aicher et al. | Substituted Tetrahydropyrrolo [2, 1-b] oxazol-5 (6 H)-ones and Tetrahydropyrrolo [2, 1-b] thiazol-5 (6 H)-ones as Hypoglycemic Agents | |
Gu et al. | Detection of endogenous hydrogen peroxide in living cells with para-nitrophenyl oxoacetyl rhodamine as turn-on mitochondria-targeted fluorescent probe | |
JP4908215B2 (en) | Treatment of disease | |
US20080027135A1 (en) | Methods For Identifying Chemical Modulators Of Ras Superfamily Gptase Activity | |
Santos et al. | Aromatic antiepileptic drugs and mitochondrial toxicity: effects on mitochondria isolated from rat liver | |
US11519893B2 (en) | Therapeutic drug for lipid-peroxidation-induced diseases and screening method for therapeutic drugs for lipid-peroxidation-induced diseases | |
Hong et al. | Isothiocyanate can be used as a highly specific recognition site for fluorescent cysteine probes | |
Zhou et al. | A sensitive pH fluorescent probe based on triethylenetetramine bearing double dansyl groups in aqueous solutions and its application in cells | |
Li et al. | A dual functional cardioinyocyte-based hybrid-biosensor for the detection of diarrhetic shellfish poisoning and paralytic shellfish poisoning toxins | |
Vhora et al. | Recent advances in in-vitro assays for type 2 diabetes mellitus: an overview | |
Gonçalves et al. | Synthesis, Characterization, and Comparative 32P-Postlabeling Efficiencies of 2, 6-Dimethylaniline− DNA Adducts | |
EP1865913A1 (en) | Sirt inhibitors that bind to nad | |
Fantinati et al. | Identification of small-molecule urea derivatives as PTPC modulators targeting the c subunit of F1/Fo-ATP synthase | |
CN104159889B (en) | Mediate noval chemical compound, the form of its mark and the application of Hedgelog protein signal path | |
CN113999159A (en) | Viscosity-sensitive fluorescent probe, and preparation method and application thereof | |
Chatterjee et al. | Interaction of tubulin with a new fluorescent analogue of vinblastine | |
Babacan et al. | Optimization of some parametric values of MTT for the determination of human melanoma (SK-Mel-30) cell viability | |
US20040110244A1 (en) | Methods for diagnosing drug-resistant cancer cells or for identifying chemotherapeutic agents by measuring anomalous intracellular ion and/or second messenger dynamics | |
Chen et al. | Mitochondrial targeted fluorescent nitrite peroxide probe for dynamic monitoring of cellular lung injury | |
CN100506769C (en) | Preparation and use of compounds i.e., 1,3-dihydroxy-5-alkyl benzene as inhibitor of protein-tyrosine-phosphatase 1B |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 20050517 |
|
AK | Designated contracting states |
Kind code of ref document: A2 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LI LU MC NL PT RO SE SI SK TR |
|
AX | Request for extension of the european patent |
Extension state: AL LT LV MK |
|
DAX | Request for extension of the european patent (deleted) | ||
RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: MILLER, GUY Inventor name: BINYAMIN, GARY Inventor name: GILAT, SYLVAIN |
|
RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: EDISON PHARMACEUTICALS, INC. |
|
A4 | Supplementary search report drawn up and despatched |
Effective date: 20080709 |
|
17Q | First examination report despatched |
Effective date: 20081111 |
|
RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: EDISON PHARMACEUTICALS, INC. |
|
RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: AMPERE LIFE SCIENCES, INC. |
|
GRAC | Information related to communication of intention to grant a patent modified |
Free format text: ORIGINAL CODE: EPIDOSCIGR1 |
|
GRAJ | Information related to disapproval of communication of intention to grant by the applicant or resumption of examination proceedings by the epo deleted |
Free format text: ORIGINAL CODE: EPIDOSDIGR1 |
|
GRAP | Despatch of communication of intention to grant a patent |
Free format text: ORIGINAL CODE: EPIDOSNIGR1 |
|
GRAC | Information related to communication of intention to grant a patent modified |
Free format text: ORIGINAL CODE: EPIDOSCIGR1 |
|
GRAC | Information related to communication of intention to grant a patent modified |
Free format text: ORIGINAL CODE: EPIDOSCIGR1 |
|
GRAP | Despatch of communication of intention to grant a patent |
Free format text: ORIGINAL CODE: EPIDOSNIGR1 |
|
GRAS | Grant fee paid |
Free format text: ORIGINAL CODE: EPIDOSNIGR3 |
|
GRAA | (expected) grant |
Free format text: ORIGINAL CODE: 0009210 |
|
AK | Designated contracting states |
Kind code of ref document: B1 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LI LU MC NL PT RO SE SI SK TR |
|
REG | Reference to a national code |
Ref country code: GB Ref legal event code: FG4D |
|
REG | Reference to a national code |
Ref country code: CH Ref legal event code: EP |
|
REG | Reference to a national code |
Ref country code: AT Ref legal event code: REF Ref document number: 598751 Country of ref document: AT Kind code of ref document: T Effective date: 20130315 |
|
REG | Reference to a national code |
Ref country code: IE Ref legal event code: FG4D |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R096 Ref document number: 60343396 Country of ref document: DE Effective date: 20130425 |
|
REG | Reference to a national code |
Ref country code: AT Ref legal event code: MK05 Ref document number: 598751 Country of ref document: AT Kind code of ref document: T Effective date: 20130227 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: SE Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 Ref country code: AT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 Ref country code: BG Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130527 Ref country code: ES Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130607 |
|
REG | Reference to a national code |
Ref country code: NL Ref legal event code: VDEP Effective date: 20130227 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: BE Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 Ref country code: FI Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 Ref country code: SI Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 Ref country code: PT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130627 Ref country code: GR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130528 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: EE Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 Ref country code: DK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 Ref country code: NL Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 Ref country code: CZ Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 Ref country code: RO Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 Ref country code: SK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: CY Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: IT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 |
|
PLBE | No opposition filed within time limit |
Free format text: ORIGINAL CODE: 0009261 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT |
|
26N | No opposition filed |
Effective date: 20131128 |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R097 Ref document number: 60343396 Country of ref document: DE Effective date: 20131128 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: MC Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 |
|
REG | Reference to a national code |
Ref country code: CH Ref legal event code: PL |
|
GBPC | Gb: european patent ceased through non-payment of renewal fee |
Effective date: 20131029 |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R119 Ref document number: 60343396 Country of ref document: DE Effective date: 20140501 |
|
REG | Reference to a national code |
Ref country code: IE Ref legal event code: MM4A |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: CH Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20131031 Ref country code: GB Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20131029 Ref country code: LI Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20131031 |
|
REG | Reference to a national code |
Ref country code: FR Ref legal event code: ST Effective date: 20140630 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: FR Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20131031 Ref country code: DE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20140501 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: IE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20131029 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: TR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20130227 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: HU Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT; INVALID AB INITIO Effective date: 20031029 Ref country code: LU Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20131029 |